cobas® MPX

Multiplex HIV, HCV & HBV nucleic acid test

for use on the cobas® 6800/8800 Systems
For in vitro diagnostic use

cobas® MPX – 96 P/N: 06997708190

cobas® MPX – 480 P/N: 06997716190

cobas® MPX Control Kit P/N: 06997724190

cobas® NHP Negative Control Kit P/N: 07002220190

cobas omni MGP Reagent P/N: 06997546190

cobas omni Specimen Diluent P/N: 06997511190

cobas omni Lysis Reagent P/N: 06997538190

cobas omni Wash Reagent P/N: 06997503190

 cobas® MPX

Table of contents
Intended use ............................................................................................................................ 4

Summary and explanation of the test................................................................................. 4

Reagents and materials ......................................................................................................... 6

cobas® MPX reagents and controls ..................................................................................................... 6

cobas omni reagents for sample preparation ..................................................................................... 10
Reagent storage and handling requirements...................................................................................... 11
Additional materials required ............................................................................................................ 12
Instrumentation and software required ............................................................................................. 12
Precautions and handling requirements ......................................................................... 13

Warnings and precautions.................................................................................................................. 13

Reagent handling ................................................................................................................................ 13
Good laboratory practice ................................................................................................................... 14
Sample collection, transport and storage ........................................................................ 15

Living donor blood samples ............................................................................................................... 15

Cadaveric blood samples .................................................................................................................... 19
Instructions for use ............................................................................................................... 20

Automated sample pipetting and pooling (optional) ....................................................................... 20

Procedural notes ................................................................................................................................. 20
Running cobas® MPX ........................................................................................................................ 20
Results ..................................................................................................................................... 21

Quality control and validity of results ................................................................................................ 21

Interpretation of results ...................................................................................................................... 22
Repeat testing of individual sample(s) ............................................................................................... 22
Procedural limitations ........................................................................................................................ 22
Non-clinical performance evaluation ............................................................................... 23

Key performance characteristics - Living donor samples .................................................................. 23

Limit of Detection (LoD).................................................................................................................................................. 23
Genotype verification ......................................................................................................................................................... 27
Seroconversion panels........................................................................................................................................................ 30
Analytical specificity............................................................................................................................................................ 34
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Whole system failure .......................................................................................................................................................... 35

Cross contamination.......................................................................................................................................................... 35
FDA/CBER panel evaluation ........................................................................................................................................... 35
Key performance characteristics - Cadaveric samples....................................................................... 38
Sensitivity............................................................................................................................................................................... 38
Specificity............................................................................................................................................................................... 38
Reproducibility .................................................................................................................................................................... 39
Clinical performance evaluation ........................................................................................ 41

Reproducibility ................................................................................................................................... 41
Clinical specificity ............................................................................................................................... 44
Reactivity in blood donor population............................................................................................................................ 44
Reactivity in source plasma donor population............................................................................................................ 44
Studies in high risk populations....................................................................................................................................... 46
Clinical sensitivity................................................................................................................................................................ 47
Studies in NAT-positive populations............................................................................................................................. 47
Clinical sensitivity for HIV-1 Group O and HIV-2 seropositive population.................................. 49
HIV-1 Group O seropositive population ..................................................................................................................... 49
HIV-2 seropositive population........................................................................................................................................ 49
Confirmation of serology results ..................................................................................................................................... 50
Additional information ......................................................................................................... 51

Key test features .................................................................................................................................. 51

Symbols ............................................................................................................................................... 52
Manufacturer and distributors .......................................................................................................... 53
Trademarks and patents ..................................................................................................................... 53
Copyright ............................................................................................................................................ 53
References ........................................................................................................................................... 54
Document revision ............................................................................................................................. 56

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Intended use
cobas® MPX, for use on cobas® 6800 and cobas® 8800 Systems is a qualitative in vitro nucleic acid screening test for the
direct detection of Human Immunodeficiency Virus Type 1 (HIV-1) Group M RNA, HIV-1 Group O RNA, Human
Immunodeficiency Virus Type 2 (HIV-2) RNA, Hepatitis C Virus (HCV) RNA, and Hepatitis B Virus (HBV) DNA in
human plasma and serum.
This test is intended for use to screen donor samples for HIV-1 Group M RNA, HIV-1 Group O RNA, HIV-2 RNA,
HCV RNA, and HBV DNA in plasma and serum samples from individual human donors, including donors of whole blood,
blood components, source plasma and other living donors. This test is also intended for use to screen organ and tissue donors
when donor samples are obtained while the donor's heart is still beating and in testing of cadaveric (non-heart beating)
donors. Plasma and serum from all donors may be screened as individual samples. For donations of whole blood and blood
components, plasma and serum samples may be tested individually or plasma may be tested in pools comprised of not more
than six individual samples. For donors of hematopoietic stem/progenitor cells (HPCs) sourced from bone marrow,
peripheral blood or cord blood, and for donors of donor lymphocytes for infusion (DLI), plasma may be tested in pools
comprised of not more than six individual samples. For donations of source plasma, samples may be tested in pools
comprised of not more than 96 individual samples. For donations from cadaveric (non-heart beating) organ and tissue
donors, samples may only be screened as individual sample.
This test is intended to be used in conjunction with licensed serology tests for HIV, HCV, and HBV.
For an individual sample, results are simultaneously detected and discriminated for HIV, HCV, and HBV.
cobas® MPX can be considered a supplemental test that confirms HIV-1 infection for samples that are repeatedly reactive
on a licensed donor screening test for antibodies to HIV-1 and reactive for HIV on cobas® MPX. cobas® MPX is not
intended to be used as a supplement test to confirm HIV-2 infection.
cobas® MPX can be considered a supplemental test that confirms HCV infection for samples that are repeatedly reactive
on a licensed donor screening test for antibodies to HCV and reactive for HCV on cobas® MPX.
cobas® MPX can be considered a supplemental test that confirms HBV infection for samples that are repeatedly reactive
on a licensed donor screening test for Hepatitis B surface antigen and reactive for HBV on cobas® MPX.
This test is not intended for use as an aid in diagnosis of HIV, HCV, or HBV.
This test is not intended for use on samples of cord blood.

Summary and explanation of the test

Background: Screening of blood for transfusion-transmitted viral infections
A major concern regarding the transfusion of blood and blood components is the potential for transmission of viral
infections, particularly with HIV-1, HIV-2, HCV, and HBV. These agents are primarily transmitted by exposure to
contaminated blood or blood and plasma products, exposure to certain body tissues or fluids, by sexual contact, or by an
infected mother to her newborn child.
HIV-1 is prevalent globally, with an estimated overall prevalence of 1.1% (0.56% in North America and 0.25% in Western
Europe).1 Persons infected with HIV-1 can experience a brief, initially acute, flu-like illness associated with high levels of
viremia in peripheral blood within 3 to 6 weeks of initial infection. There are currently three principal genetic groups for
HIV-1: Group M (main), Group N (non-M-non-O), and Group O (outlier). Group M is highly prevalent and is divided
into 9 subtypes, as well as several circulating recombinant forms (CRFs).2-4
HIV-2 was first isolated in 1986 from patients in West Africa. Both HIV-1 and HIV-2 have the same modes of
transmission and are associated with similar opportunistic infections and Acquired Immunodeficiency Syndrome
(AIDS).5, 6 The prevalence of HIV-2 in some African nations reaches more than 1%, and HIV-2 is a growing concern in
certain parts of Europe and India.7-11 The first case of HIV-2 infection in the United States (US) was diagnosed in 1987.
The Centers for Disease Control and Prevention (CDC) advise that continued surveillance is needed to monitor HIV-2 in
the US population.12
HCV is considered to be the principal etiologic agent responsible for 90% to 95% of post transfusion non-A and non-B hepatitis
cases.12-15 The reported prevalence of HCV varies from 0.5 to 2.0% in Western Europe16 and between 6% and 40% in Egypt.17
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More than 2 billion people alive today have been infected with HBV at some time in their lives. Of these, about 350 million
remain infected chronically and become carriers of the virus.18-20 Both HCV and HBV can result in chronic liver disease, and
these viruses are the most common cause of liver cirrhosis and cancer, accounting for 78% of cases globally.21
Rationale for NAT testing
Serological screening assays have greatly reduced, but not eliminated, the risk of transmission of viral infections by
transfusion of blood and blood products. Testing of whole blood and source plasma donations for HBV was initiated with
HBsAg assays in the early 1970s and anti-HBc in the 1980s. In addition to HBV screening, blood and plasma donations are
routinely tested for antibodies to HIV and HCV using enzyme immunoassays (EIAs).22, 23 A residual transmission risk
exists from blood donations made during the seroconversion window period, which has been estimated to be
approximately 19 days, 65 days and 36 days for HIV-1, HCV and HBV, respectively.24 Testing for the viral nucleic acids
(HIV-1 RNA, HCV RNA, and HBV DNA), using nucleic acid amplification technology (NAT) can substantially reduce
this risk.25,26 With the introduction of NAT, the current residual risk of transfusion in the US is 1:1.5 million for HIV-1,
1:1.2 million for HCV and 1:280,000-1:355,000 for HBV.27,28 Similar estimates for Germany, where NAT testing was
introduced in 1999, give an estimated residual risk of transfusion transmitted infections of 1:4.3 million, 1:10.9 million and
1:360,000, for HIV-1, HCV and HBV respectively. 25 In addition, in the case of HBV, NAT testing will also interdict donors
with an occult HBV infection in which HBV DNA is detectable but HBsAg is absent,29 and in vaccinated donors with a
breakthrough, subclinical infection.30-32
Explanation of the test
cobas® MPX is a qualitative multiplex test that is run on the cobas® 6800 System and cobas® 8800 System. cobas® MPX
enables the simultaneous detection and discrimination of HIV RNA, HCV RNA, HBV DNA, and the internal control in a
single test of an infected, individual donation or pooled plasma from individual donations. The test does not discriminate
between HIV-1 Group M, HIV-1 Group O, and HIV-2.
Principles of the procedure
cobas® MPX is based on real time PCR technology on a fully automated sample preparation (nucleic acid extraction and
purification) followed by PCR amplification and detection system. The cobas® 6800/8800 Systems consist of the sample
supply module, the transfer module, the processing module, and the analytic module. Automated data management is
performed by the cobas® 6800/8800 software which assigns test results for all tests as non-reactive, reactive, or invalid.
Results can be reviewed directly on the system screen, and printed as a report , or sent to a Laboratory Information
Management System (LIMS) or other result management system.
Samples can either be tested individually or, optionally, can be tested in pools consisting of multiple samples. The
cobas p 680 instrument may optionally be used in a pre-analytical step if pooling is to be performed.
Nucleic acid from the sample and added armored RNA internal control (IC) molecules (which serve as the sample
preparation and amplification/detection process control) is simultaneously extracted. In addition the test utilizes four
external controls: three positive and a negative control. Viral nucleic acid is released by addition of proteinase and lysis
reagent to the sample. The released nucleic acid binds to the silica surface of the added magnetic glass particles. Unbound
substances and impurities, such as denatured protein, cellular debris, and potential PCR inhibitors (such as hemoglobin)
are removed with subsequent wash reagent steps and purified nucleic acid is eluted from the magnetic glass particles with
elution buffer at elevated temperature.
Selective amplification of target nucleic acid from the donor sample is achieved by the use of virus-specific forward and
reverse primers which are selected from highly conserved regions of the viral nucleic acid. A thermostable DNA polymerase
enzyme is used for both reverse-transcription and amplification. The master mix includes deoxyuridine triphosphate
(dUTP), instead of deoxythimidine triphosphate (dTTP), which is incorporated into the newly synthesized DNA
(amplicon).33-35 Any contaminating amplicon from previous PCR runs are eliminated by the AmpErase enzyme [uracil-
N-glycosylase], which is included in the PCR master mix, when heated in the first thermal cycling step. However, newly
formed amplicon are not eliminated since the AmpErase enzyme is inactivated once exposed to temperatures above 55°C.
The cobas® MPX master mix contains detection probes which are specific for HIV-1 (Groups M and O), HIV-2, HCV,
HBV, and IC nucleic acid. The specific HIV, HCV, HBV, and IC detection probes are each labeled with one of four
unique fluorescent dyes which act as a reporter. Each probe also has a fifth dye which acts as a quencher. The four
reporter dyes are measured at defined wavelengths, thus permitting simultaneous detection and discrimination of the
amplified HIV, HCV, and HBV targets and the IC. 36, 37 When not bound to the target sequence, the fluorescent signal of
the intact probes is suppressed by the quencher dye. During the PCR amplification step, hybridization of the probes to

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the specific single-stranded DNA template results in cleavage by the 5' to 3' nuclease activity of the DNA polymerase
resulting in separation of the reporter and quencher dyes and the generation of a fluorescent signal. With each PCR cycle,
increasing amounts of cleaved probes are generated and the cumulative signal of the reporter dye is concomitantly increased.
Since the four specific reporter dyes are measured at defined wavelengths, simultaneous detection and discrimination of the
amplified HIV, HCV and HBV targets and the IC are possible.

Reagents and materials

cobas® MPX reagents and controls
All unopened reagents and controls shall be stored as recommended in Table 1 to Table 4.

Table 1 cobas® MPX

cobas® MPX
Store at 2-8°C
96 test cassette (P/N 06997708190)
480 test cassette (P/N 06997716190)
Kit components Reagent ingredients Quantity per kit
96 tests 480 tests

Proteinase Solution Tris buffer, < 0.05% EDTA, calcium chloride, calcium acetate, 13 mL 38 mL
(PASE) 8% (w/v) proteinase
EUH210: Safety data sheets available on request.
EUH208: Contains: Subtilisin, 9014-01-1. May produce an
allergic reaction.
Internal Control Tris buffer, < 0.05% EDTA, < 0.001% internal control armored 13 mL 38 mL
(IC) RNA construct (non-infectious RNA encapsulated in MS2
bacteriophage), < 0.002% Poly rA RNA (synthetic), < 0.1%
sodium azide
Elution Buffer Tris buffer, 0.2% methyl-4 hydroxybenzoate 13 mL 38 mL
(EB)
Master Mix Manganese acetate, potassium hydroxide, < 0.1% sodium 5.5 mL 14.5 mL
Reagent 1 azide
(MMX-R1)
MPX Master Mix Tricine buffer, potassium acetate, glycerol, 18% dimethyl 6 mL 17.5 mL
Reagent 2 sulfoxide, Tween 20, EDTA, < 0.06% dATP, dGTP, dCTP,
(MPX MMX-R2) < 0.14% dUTP, < 0.01% upstream and downstream HIV-1
Group M, HIV-1 Group O, HIV-2, HCV, HBV, and internal
control primers, < 0.01% fluorescent-labeled HIV, HCV, and
HBV probes, < 0.01% fluorescent-labeled internal control probe,
< 0.01% oligonucleotide aptamer, < 0.01% ZO5D DNA
polymerase, < 0.01% AmpErase (uracil-N-glycosylase) enzyme
(microbial), < 0.1% sodium azide

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Table 2 cobas® MPX Control Kit

cobas® MPX Control Kit

Store at 2-8°C
(P/N 06997724190)
Kit components Reagent ingredients Quantity Safety symbol and warning*
per kit
MPX Multi- < 0.001% Synthetic (armored) HIV-1 Group M 4 mL
Positive Control RNA encapsulated in MS2 bacteriophage coat (4 x 1 mL)
(MPX M (+) C) protein, < 0.001% synthetic (armored) HCV RNA
encapsulated in MS2 bacteriophage coat
protein, < 0.001% Synthetic (plasmid) HBV DNA
encapsulated in Lambda bacteriophage coat Warning
protein, normal human plasma, non-reactive by H317: May cause an allergic skin reaction.
licensed tests for antibody to HCV, antibody to P261: Avoid breathing
HIV-1/2, HBsAg, antibody to HBc; HIV-1 RNA, dust/fumes/gas/mist/vapors/spray.
HIV-2 RNA, HCV RNA, HBV DNA, HEV RNA,
P272: Contaminated work clothing should
WNV RNA, and CMV DNA not detectable by
not be allowed out of the workplace.
PCR methods.
P280: Wear protective gloves.
0.1% ProClin® 300 preservative
P302+ P352: IF ON SKIN wash with plenty
of soap and water.
P333 + P313: If skin irritation or rash
occurs: Get medical advice/attention.
P363: Wash contaminated clothing before
reuse.
MPX HIV-1 O < 0.001% Synthetic (armored) HIV-1 Group O 4 mL
Positive Control RNA encapsulated in MS2 bacteriophage coat (4 x 1 mL)
(MPX O (+) C) protein, normal human plasma, non-reactive by
licensed tests for antibody to HCV, antibody to
HIV-1/2, HBsAg, antibody to HBc; HIV-1 RNA,
HIV-2 RNA, HCV RNA, HBV DNA, HEV RNA, Warning
WNV RNA, and CMV DNA not detectable by H317: May cause an allergic skin reaction.
PCR methods. P261: Avoid breathing
®
0.1% ProClin 300 preservative dust/fumes/gas/mist/vapors/spray.
P272: Contaminated work clothing should
not be allowed out of the workplace.
P280: Wear protective gloves.
P302+ P352: IF ON SKIN wash with plenty
of soap and water.
P333 + P313: If skin irritation or rash
occurs: Get medical advice/attention.
P363: Wash contaminated clothing before
reuse.

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Table 2 cobas® MPX Control Kit

cobas® MPX Control Kit

Store at 2-8°C
(P/N 06997724190)
Kit components Reagent ingredients Quantity Safety symbol and warning*
per kit
MPX HIV-2 < 0.001% Synthetic (armored) HIV-2 RNA 4 mL
Positive Control encapsulated in MS2 bacteriophage coat (4 x 1 mL)
(MPX 2 (+) C) protein, normal human plasma, non-reactive by
licensed tests for antibody to HCV, antibody to
HIV-1/2, HBsAg, antibody to HBc; HIV-1 RNA,
HIV-2 RNA, HCV RNA, HBV DNA, HEV RNA, Warning
WNV RNA, and CMV DNA not detectable by H317: May cause an allergic skin reaction.
PCR methods. P261: Avoid breathing
®
0.1% ProClin 300 preservative dust/fumes/gas/mist/vapors/spray.
P272: Contaminated work clothing should
not be allowed out of the workplace.
P280: Wear protective gloves.
P302+ P352: IF ON SKIN wash with plenty
of soap and water.
P333 + P313: If skin irritation or rash
occurs: Get medical advice/attention.
P363: Wash contaminated clothing before
reuse.
* Product safety labeling primarily follows EU GHS guidance

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Table 3 cobas® NHP Negative Control Kit

cobas® NHP Negative Control Kit

Store at 2-8°C
(P/N 07002220190)
Kit components Reagent ingredients Quantity Safety symbol and warning*
per kit
Normal Human Normal human plasma, non-reactive by 16 mL
Plasma Negative licensed tests for antibody to HCV, antibody (16 x 1 mL)
Control to HIV-1/2, HBsAg, antibody to HBc; HIV-1
(NHP-NC) RNA, HIV-2 RNA, HCV RNA, HBV DNA, HEV
RNA, WNV RNA, and CMV DNA not
detectable by PCR methods. Warning
H317: May cause an allergic skin reaction.
< 0.1% ProClin® 300 preservative
P261: Avoid breathing dust/fumes/gas/mist/
vapors/spray.
P272: Contaminated work clothing should
not be allowed out of the workplace.
P280: Wear protective gloves.
P302+ P352: IF ON SKIN wash with plenty
of soap and water.
P333 + P313: If skin irritation or rash occurs:
Get medical advice/attention.
P363: Wash contaminated clothing before
reuse.

* Product safety labeling primarily follows EU GHS guidance

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cobas omni reagents for sample preparation

Table 4 cobas omni reagents for sample preparation*

Reagents Reagent ingredients Quantity Safety symbol and warning**

per kit
cobas omni Magnetic glass particles, Tris buffer, 0.1% 480 tests Not applicable
MGP Reagent methyl-4 hydroxybenzoate, < 0.1% sodium
(MGP) azide
Store at 2–8°C
(P/N 06997546190)
cobas omni Tris buffer, 0.1% methyl-4 4 x 875 mL Not applicable
Specimen Diluent hydroxybenzoate, < 0.1% sodium azide
(SPEC DIL)
Store at 2–8°C
(P/N 06997511190)
cobas omni 42.56% (w/w) guanidine thiocyanate, 5% 4 x 875 mL
Lysis Reagent (w/v) polydocanol, 2% (w/v) dithiothreitol,
(LYS) dihydro sodium citrate
Store at 2–8°C
(P/N 06997538190) Danger
H302: Harmful if swallowed.
H318: Causes serious eye damage.
H412: Harmful to aquatic life with long lasting effects.
EUH032: Contact with acids liberates very toxic gas.
P264: Wash skin thoroughly after handling.
P270: Do not eat, drink or smoke when using this product.
P273: Avoid release to the environment.
P280: Wear protective gloves/eye protection/face
protection.
P301 + P312: IF SWALLOWED: Call a POISON CENTER or
doctor/physician if you feel unwell.
P305+P351+P338: IF IN EYES Rinse cautiously with water
for several minutes. Remove contact lenses, if present and
easy to do. Continue rinsing.
P310: Immediately call a POISON CENTER or
doctor/physician if you feel unwell.
P330: Rinse mouth.
cobas omni Sodium citrate dihydrate, 0.1% methyl-4 4.2 L Not applicable
Wash Reagent hydroxybenzoate
(WASH)
Store at 15–30°C
(P/N 06997503190)

* These reagents are not included in the cobas® MPX test kit. See listing of additional materials required (Table 7).
**Product safety labeling primarily follows EU GHS guidance

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Reagent storage and handling requirements

Opened reagents shall be stored and will be handled as specified in Table 5 and Table 6.
When reagents are not loaded on the cobas® 6800/8800 Systems, store them at the corresponding temperature specified in
Table 5.
Table 5 Reagent storage (when reagent is not on the system)

Reagent Storage temperature

cobas® MPX – 96 2–8°C
cobas® MPX – 480 2–8°C
cobas® MPX Control Kit 2–8°C
cobas® NHP Negative Control Kit 2–8°C
cobas omni Lysis Reagent 2–8°C
cobas omni MGP Reagent 2–8°C
cobas omni Specimen Diluent 2–8°C
cobas omni Wash Reagent 15–30°C

Reagents loaded onto the cobas® 6800/8800 Systems are stored at appropriate temperatures and their expiration is
monitored by the system. The system allows reagents to be used only if all of the conditions shown in Table 6 are met. The
system automatically prevents use of expired reagents. Table 6 allows the user to understand the reagent handling
conditions enforced by the cobas® 6800/8800 Systems.
Table 6 Reagent expiry conditions enforced by the cobas® 6800/8800 Systems

Reagent Kit expiration Open-kit stability Number of runs for On-board stability
date which this kit can (cumulative time on
be used board outside
refrigerator)

cobas® MPX – 96 Date not passed 30 days from first usage Max 10 runs Max 8 hours

cobas® MPX – 480 Date not passed 30 days from first usage Max 20 runs Max 20 hours

cobas® MPX Control Kit Date not passed Not applicable Not applicable Max 8 hours

cobas® NHP Negative Control Kit Date not passed Not applicable Not applicable Max 10 hours

cobas omni Lysis Reagent Date not passed 30 days from loading* Not applicable Not applicable

cobas omni MGP Reagent Date not passed 30 days from loading* Not applicable Not applicable

cobas omni Specimen Diluent Date not passed 30 days from loading* Not applicable Not applicable

cobas omni Wash Reagent Date not passed 30 days from loading* Not applicable Not applicable

* Time is measured from the first time that reagent is loaded onto the cobas® 6800/8800 Systems.

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Additional materials required

Table 7 Material and consumables for use on cobas® 6800/8800 Systems

Material P/N

cobas omni Processing Plate 05534917001

cobas omni Amplification Plate 05534941001

cobas omni Pipette Tips 05534925001

cobas omni Liquid Waste Container 07094388001

cobas omni Lysis Reagent 06997538190

cobas omni MGP Reagent 06997546190

cobas omni Specimen Diluent 06997511190

cobas omni Wash Reagent 06997503190

Solid Waste Bag 07435967001

Solid Waste Container 07094361001

Instrumentation and software required

The cobas® 6800/8800 software and cobas® MPX analysis package shall be installed on the instrument(s). The Instrument
Gateway (IG) server will be provided with the system.

Table 8 Instrumentation

Equipment P/N

cobas® 6800 System (Option Moveable) 05524245001 and 06379672001

cobas® 6800 System (Fix) 05524245001 and 06379664001

cobas® 8800 System 05412722001

cobas p 680 Instrument (optional for pipetting and pooling) 06570577001

Sample Supply Module 06301037001

Refer to the cobas® 6800/8800 Systems Operator’s Manual and cobas p 680 instrument Operator’s Manual for additional information
for primary and secondary sample tubes accepted on the instruments.
Note: Contact your local Roche representative for a detailed order list for sample racks, racks for clotted tips and rack trays accepted
on the instruments.

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Precautions and handling requirements

Warnings and precautions
As with any test procedure, good laboratory practice is essential to the proper performance of this assay. Due to the high
sensitivity of this test, care should be taken to keep reagents and amplification mixtures free of contamination.
 For in vitro diagnostic use only.
 All samples should be handled as if infectious, using good laboratory procedures as outlined in Biosafety in
Microbiological and Biomedical Laboratories and in the CLSI Document M29-A4.38,39 Only personnel proficient in
handling infectious materials and the use of cobas® MPX, cobas® 6800/8800 Systems and cobas p 680 instrument
(optional) should perform this procedure.
 All human-sourced materials should be considered potentially infectious and should be handled with universal
precautions. If spillage occurs, immediately disinfect with a freshly prepared solution of 0.5% sodium hypochlorite
in distilled or deionized water (dilute household bleach 1:10) or follow appropriate site procedures.
 cobas® MPX Control Kit and cobas® NHP Negative Control Kit contain plasma derived from human blood. The
source material has been tested by licensed antibody tests and found non-reactive for the presence of antibody to
HCV, antibody to HIV-1/2, HBsAg, and antibody to HBc. Testing of normal human plasma by PCR methods also
showed no detectable HIV-1 (Groups M and O) RNA, HIV-2 RNA, HCV RNA, HBV DNA, HEV RNA, WNV
RNA, or CMV DNA. No known test method can offer complete assurance that products derived from human blood
will not transmit infectious agents.
 Do not freeze whole blood.
 The use of sterile disposable pipettes and nuclease-free pipette tips is recommended. Use only supplied or specified
required consumables to ensure optimal test performance.
 Closely follow procedures and guidelines provided to ensure that the test is performed correctly. Any deviation
from the procedures and guidelines may affect optimal test performance.
 False positive results may occur if carryover of samples is not adequately controlled during sample handling
and processing.

Reagent handling
 Handle all reagents, controls, and samples according to good laboratory practice in order to prevent carryover of
samples or controls.
 Before use, visually inspect each reagent cassette, diluent, lysis reagent, and wash reagent to ensure that there are no
signs of leakage. If there is any evidence of leakage, do not use that material for testing.
 cobas omni Lysis Reagent contains guanidine thiocyanate, a potentially hazardous chemical. Avoid contact of
reagents with the skin, eyes, or mucous membranes. If contact does occur, immediately wash with generous
amounts of water; otherwise, burns can occur.
 cobas® MPX kits, cobas omni MGP Reagent, and cobas omni Specimen Diluent contain sodium azide as a
preservative. Avoid contact of reagents with the skin, eyes, or mucous membranes. If contact does occur,
immediately wash with generous amounts of water; otherwise, burns can occur. If these reagents are spilled, dilute
with water before wiping dry.
 Do not allow cobas omni Lysis Reagent, which contains guanidine thiocyanate, to contact sodium hypochlorite
(bleach) solution. This mixture can produce a highly toxic gas.
 Safety Data Sheets (SDS) are available on request from your local Roche representative.
 Dispose of all materials that have come in contact with samples and reagents in accordance with country, state, and
local regulations.

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Good laboratory practice

 Do not pipette by mouth.
 Do not eat, drink, or smoke in designated work areas.
 Wear laboratory gloves, laboratory coats, and eye protection when handling samples and reagents. Gloves must be
changed between handling samples and cobas® MPX kits and cobas omni reagents to prevent contamination.
Avoid contaminating gloves when handling samples and controls.
 Wash hands thoroughly after handling samples and kit reagents, and after removing the gloves.
 Thoroughly clean and disinfect all laboratory work surfaces with a freshly prepared solution of 0.5% sodium
hypochlorite in distilled or deionized water (dilute household bleach 1:10). Follow by wiping the surface with
70% ethanol.
 If spills occur on the cobas® 6800/8800 instrument, follow the instructions in the cobas® 6800/8800 System
Operator’s Manual to properly clean and decontaminate the surface of instrument(s).

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Sample collection, transport and storage

Note: Handle all samples and controls as if they are capable of transmitting infectious agents.
Store all donor samples at specified temperatures.
Sample stability is affected by elevated temperatures.

 It is recommended that serum samples are tested within 8 hours of centrifugation at 1600 x g for 20 minutes or are
tested within 24 hours of high-speed centrifugation (e.g., 2600 x g for 20 minutes).

Living donor blood samples

 Plasma collected in EDTA, CPD, CPDA1, CP2D and 4% sodium citrate anticoagulant and serum collected in
serum clot tubes may be used with cobas® MPX. Follow the sample collection tube/bag manufacturer instructions
for handling and centrifugation.
 Blood collected in EDTA anticoagulant may be stored for up to 12 days with the following conditions:
o Samples must be centrifuged within 72 hours of draw.
o For storage above 8°C, samples may be stored for 72 hours at up to 25°C, and up to 30°C for 24 hours
during the 72 hours.
Other than noted above, samples are stored at 2-8°C. In addition, plasma separated from the cells may be stored for up to
12 months at <-18°C with three freeze/thaw cycles. Refer to Figure 1.

Figure 1 Sample storage conditions for living donor sample in EDTA anticoagulant

30
EDTA anticoagulant

25
Temperature °C

8
Whole Blood
2
1 3 12
≤-18
Frozen plasma
(Freeze-thaw up to 3 times)

Days 0 Months 12

 Blood collected in CPD, CPDA1, CP2D anticoagulant, Becton-Dickinson EDTA Plasma Preparation Tubes (BD PPT™)
or Greiner Vacuette® K2EDTA Plasma Gel Tubes may be stored for up to 12 days with the following conditions:
o Samples must be centrifuged within 72 hours of draw.
o For storage above 8°C, samples may be stored for 72 hours at up to 25°C, and up to 30°C for 24 hours
during the 72 hours.
Other than noted above, samples are stored at 2-8°C. In addition, plasma separated from the cells may be stored for up to
30 days at <-18°C with three freeze/thaw cycles. Refer to Figure 2.

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Figure 2 Sample storage conditions for living donor sample

TM ®
CPD, CPDA1, CP2D anticoagulant, BD PPT or Greiner Vacuette K2EDTA
30
Plasma Gel Tubes

25
Temperature °C

8
Whole Blood

2
1 3 12
≤-18
Frozen plasma
18 (Freeze-thaw up to 3 times)

0 30
Days

 Blood collected in serum clot tubes may be stored for up to 7 days at 2-8°C with the following conditions,
o Samples must be centrifuged within 72 hours of draw.
o For storage above 8°C, samples may be stored for 72 hours at up to 25°C, and up to 30°C for 24 hours
during the 72 hours.
Other than noted above, samples are stored at 2-8°C. In addition, serum separated from the cells may be stored for up to
30 days at ≤ -18°C with three freeze/thaw cycles.
 Plasma collected in 4% sodium citrate anticoagulant may be stored for up to 30 days at 2-8°C with the following
two conditions:
o For storage above 8°C, specimens may be stored for 72 hours at up to 25°C, and up to 30°C for 24 hours
during the 72 hours.
In addition, plasma collected in 4% sodium citrate anticoagulant may be stored for up to 12 months at ≤ -18°C
with two freeze/thaw cycles. Refer to Figure 3.

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Figure 3 Sample storage conditions for plasma

30
4% Sodium Citrate

25
Temperature °C

8
Plasma
2
1 3 30
≤-18
Frozen plasma
(Freeze-thaw up to 2 times)

Days 0 12
Months

 Plasma collected in 4% sodium citrate anticoagulant may be stored for up to 18 days at 2-8°C with the following
conditions:
o For storage above 8°C, samples may be stored for 72 hours at up to 25°C, and up to 30°C for 24 hours
during the 72 hours.
In addition, apheresis plasma collected in 4% sodium citrate anticoagulant may be stored for up to 12 months
at ≤ - 18°C with three freeze/thaw cycles. Refer to Figure 4.

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Figure 4 Sample storage conditions for plasma

30
4% Sodium Citrate

25
Temperature °C

8
Plasma
2
1 3 18
≤-18
Frozen plasma
(Freeze-thaw up to 3 times)

0 12
Days Months

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Cadaveric blood samples

 Cadaveric blood samples collected in EDTA anticoagulant tubes and/or in serum clot tubes may be used with the
cobas® MPX test. Follow the sample collection tube/bag manufacturer instructions for handling and
centrifugation.
 Cadaveric blood collected in EDTA anticoagulant may be stored for up to 8 days at 2-8°C with the following
conditions:
o Samples must be centrifuged within 72 hours of draw.
o For storage above 8°C, samples may be stored at up to 30°C, for 24 hours during the 72 hours.
Other than noted above, cadaveric EDTA plasma separated from the cells may be stored for up to 14 days at ≤-18°C.
Refer to Figure 5.

Figure 5 Sample storage conditions for cadaveric sample

30
EDTA anticoagulant

25
Temperature °C

8
Whole Blood

2
1 8
≤-18
Frozen plasma

0 14
Days

 Cadaveric blood samples collected in serum clot tubes may be stored for up to 5 days at 2-8°C with the following
conditions:
o Samples must be centrifuged within 72 hours of draw.
o For storage above 8°C, samples may be stored for 24 hours at up to 30°C, during the 72 hours.
 If living donor and/or cadaveric samples are to be shipped, they should be packaged and labeled in compliance
with applicable country and/or international regulations covering the transport of samples and etiologic agents.

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Instructions for use

Automated sample pipetting and pooling (optional)
The cobas p 680 instrument is an optional component of the cobas® 6800/8800 Systems used for automated pipetting and
pooling of aliquots of multiple primary samples into one pooled sample. Refer to the cobas p 680 instrument Operator’s
Manual for more information.

Procedural notes
 Do not use cobas® MPX reagents, cobas® MPX Control Kit, cobas® NHP Negative Control Kit, or cobas omni
reagents after their expiry dates.
 Do not reuse consumables. They are for one-time use only.
 Refer to the cobas® 6800/8800 Systems Operator’s Manual for proper maintenance of instruments.

Running cobas® MPX

The test procedure is described in detail in the cobas® 6800/8800 Systems Operator’s Manual and the cobas p 680
instrument Operator’s Manual. Figure 6 below summarizes the procedure.
®
Figure 6 cobas MPX procedure

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Results
The cobas® 6800/8800 System automatically detects and discriminates HIV RNA, HCV RNA, and HBV DNA
simultaneously for the samples and controls.

Quality control and validity of results

 One negative control [(–) C] and three positive controls [MPX M (+) C, MPX O (+) C, and MPX 2 (+) C] are
processed with each batch.
 In the cobas® 6800/8800 software and/or report, check for flags and their associated results to ensure the batch validity.
 The batch is valid if no flags appear for all four controls.
Invalidation of results is performed automatically by the cobas® 6800/8800 software based on negative and positive
control failures.

Control flags

Table 9 Control flags for negative and positive controls

Negative Control Flag Result Interpretation

(-) C Q02 Invalid The entire batch is assigned invalid if the result for the (-) C is invalid.
Positive Control Flag Result Interpretation
MPX M (+) C Q02 Invalid The entire batch is assigned invalid if the result for the MPX M (+) C is invalid.
MPX O (+) C Q02 Invalid The entire batch is assigned invalid if the result for the MPX O (+) C is invalid.
MPX 2 (+) C Q02 Invalid The entire batch is assigned invalid if the result for the MPX 2 (+) C is invalid.

If the batch is invalid, repeat testing of the entire batch including samples and controls.

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Interpretation of results
For a valid batch, check each individual sample for flags in the cobas® 6800/8800 software and/or report. The result
interpretation should be as follows:
 A valid batch may include both valid and invalid donor sample results dependent on flags obtained for the
individual samples.
 Sample results are valid only if the respective positive controls and the negative control of the corresponding batch
are valid.
Four parameters are measured simultaneously for each sample: HIV, HCV, HBV, and the internal control. Final sample
results for cobas® MPX are reported by the software. In addition to the overall results, individual target results will be
displayed in the cobas® 6800/8800 software and should be interpreted as follows:

Table 10 Target results for individual target result interpretation

Target results Interpretation

HIV Non-Reactive No target signal detected for HIV and IC signal detected.
HIV Reactive Target signal detected for HIV and IC signal may be or may not be detected.
HCV Non-Reactive No target signal detected for HCV and IC signal detected.
HCV Reactive Target signal detected for HCV and IC signal may be or may not be detected.
HBV Non-Reactive No target signal detected for HBV and IC signal detected.
HBV Reactive Target signal detected for HBV and IC signal may be or may not be detected.
Invalid Target and internal control signal not detected.

Repeat testing of individual sample(s)

Sample tubes with a final result of Invalid for one target require repeat testing regardless of valid results for the other targets.

Procedural limitations
 cobas® MPX has been evaluated only for use in combination with the cobas® MPX Control Kit, cobas® NHP
Negative Control Kit, cobas omni MGP Reagent, cobas omni Lysis Reagent, cobas omni Specimen Diluent, and
cobas omni Wash Reagent for use on the cobas® 6800/8800 Systems.
 Reliable results depend on proper sample collection, storage and handling procedures.
 Do not use heparinized plasma with this test because heparin has been shown to inhibit PCR.
 Detection of HIV-1 Group M RNA, HIV-1 Group O RNA, HIV-2 RNA, HCV RNA, and HBV DNA is dependent
on the number of virus particles present in the sample and may be affected by sample collection, storage and
handling, patient factors (i.e., age, presence of symptoms), and/or stage of infection and pool size.
 Though rare, mutations within the highly conserved regions of a viral genome covered by cobas® MPX, may affect
primers and/or probe binding resulting in the failure to detect presence of virus.
 Due to inherent differences between technologies, it is recommended that, prior to switching from one technology
to the next, users perform method correlation studies in their laboratory to qualify technology differences. Users
should follow their own specific policies/procedures.

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Non-clinical performance evaluation

Key performance characteristics - Living donor samples

Limit of Detection (LoD)

WHO International Standards/Roche Primary Standards

The limits of detection (LoD) of cobas® MPX for HIV-1 Group M RNA HIV-1 Group O RNA, HIV-2 RNA, HCV RNA,
and HBV DNA were determined using the following standards:
 WHO 3rd International Standard for HIV-1 Group M RNA (NIBSC code 10/152)
 WHO International Standard for HIV-2 RNA (NIBSC code 08/150)40
 Roche Primary Standards for HIV-1 Group O RNA
 WHO 2nd International Standard for HCV RNA (NIBSC code 96/798)
 WHO 3rd International Standard for HBV DNA (NIBSC 10/264)
No international standard is currently available for HIV-1 Group O RNA. The Roche HIV-1 Group O RNA Standard is
traceable to the CBER HIV-1 Subtype RNA Reference Panel #1 Lot 01. The Roche Primary Standards for HIV-1 Group O
RNA are derived from commercially available cultured virus stocks, P/N 2420 (Cat. No. 500493, SeraCare Life Sciences).
For the WHO International HIV-1 Group M, HCV and HBV, HIV-2, and Roche primary HIV-1 Group O standards, three
independent dilution series of each viral standard co-formulated for HIV-1 Group M, HCV, and HBV members and
individually formulated HIV-1 Group O, and HIV-2 were prepared with normal, virus-negative (HIV, HBV and HCV)
human EDTA-plasma. Each dilution series was tested using three different lots of cobas® MPX kits with approximately
63 replicates per lot, for a total of approximately 189 replicates per concentration. For the WHO International HIV-2
Standard, 33 replicates per lot from three independent dilutions and three reagent lots were tested for a total of
99 replicates per concentration. For each virus, PROBIT analysis on the data combined across dilution series and reagent
lots was used to estimate the LoD, along with the lower and upper limit of the 95% confidence interval (Table 11). The
reactivity rates observed in the LoD studies for each virus are summarized in Table 12 to Table 16.

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Table 11 Results of PROBIT analysis on LoD data collected with viral standards in EDTA plasma and serum

Measuring Lower 95% Upper 95%

Matrices Analyte LoD
units confidence limit confidence limit

HIV-1 Group M IU/mL 25.7 21.1 32.8

HIV-1 Group O copies/mL 8.2 7.0 10.0
EDTA Plasma HIV-2 IU/mL 4.0 3.3 5.2
HCV IU/mL 7.0 5.9 8.6
HBV IU/mL 1.4 1.2 1.7
HIV-1 Group M IU/mL 23.7 20.0 29.1
HIV-1 Group O copies/mL 12.2 10.3 14.9
Serum HIV-2 IU/mL 4.4 3.5 5.8
HCV IU/mL 8.1 6.8 10.1
HBV IU/mL 1.3 1.1 1.5

Table 12 Reactivity rates summary for HIV-1 Group M in EDTA plasma and serum

HIV-1 Group M RNA 95% Lower

Number of valid
Matrices concentration Number reactive % Reactive confidence bound
replicates
(IU/mL) (one-sided)
30 186 188 98.9% 96.7%
15 170 189 89.9% 85.6%
EDTA Plasma 7.5 124 189 65.6% 59.5%
4.5 96 189 50.8% 44.6%
1.5 50 189 26.5% 21.2%
30 186 189 98.4% 95.9%
15 170 189 89.9 % 85.6%
Serum 7.5 123 189 65.1% 59.0%
4.5 85 189 45.0% 38.8%
1.5 31 189 16.4% 12.1%

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Table 13 Reactivity rates summary for HIV-1 Group O in EDTA plasma and serum

HIV-1 Group O RNA 95% Lower

Number of valid
Matrices concentration Number reactive % Reactive confidence bound
replicates
(copies/mL) (one-sided)
18 187 187 100.0% 98.4%
9 181 187 96.8% 93.8%
EDTA Plasma 4.5 162 189 85.7% 80.8%
2.7 117 189 61.9% 55.7%
0.9 57 189 30.2% 24.7%
18 186 187 99.5% 97.5%
9 173 188 92.0% 88.0%
Serum 4.5 142 189 75.1% 69.4%
2.7 79 189 41.8% 35.8%
0.9 39 189 20.6% 15.9%

Table 14 Reactivity rates summary for HIV-2 in EDTA plasma and serum

HIV-2 95% Lower

Number of valid
Matrices RNA concentration Number reactive % Reactive confidence bound
replicates
(IU/mL) (one-sided)

10 98 98 100.0% 97.0%
5 98 99 99.0% 95.3 %
EDTA Plasma 2.5 80 98 81.6% 74.0%
1.5 71 99 71.7% 63.3%
0.5 26 99 26.3% 19.1%
10 98 98 100.0% 97.0%
5 98 99 99.0% 95.3%
Serum 2.5 81 99 81.8% 74.2%
1.5 63 98 64.3% 55.6%
0.5 28 98 28.6% 21.1%

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Table 15 Reactivity rates summary for HCV in EDTA plasma and serum

HCV RNA 95% Lower

Number of valid
Matrices concentration Number reactive % Reactive confidence bound
replicates
(IU/mL) (one-sided)
12 187 188 99.5 % 97.5 %
6 178 189 94.2 % 90.6 %
EDTA Plasma 3 148 189 78.3 % 72.8 %
1.8 112 189 59.3 % 53.0 %
0.6 50 189 26.5 % 21.2 %
12 186 189 98.4 % 95.9 %
6 173 189 91.5 % 87.4 %
Serum 3 139 189 73.5 % 67.7 %
1.8 112 189 59.3 % 53.0 %
0.6 41 189 21.7 % 16.9 %

Table 16 Reactivity rates summary for HBV in EDTA plasma and serum

HBV 95% Lower

Number of valid
Matrices DNA concentration Number reactive % Reactive confidence bound
replicates
(IU/mL) (one-sided)

3.40 188 188 100.0 % 98.4 %

1.70 184 189 97.4 % 94.5 %

EDTA plasma 0.85 165 189 87.3 % 82.6 %

0.51 126 189 66.7 % 60.6 %

0.17 58 189 30.7 % 25.2 %

3.40 189 189 100.0 % 98.4 %

1.70 184 189 97.4 % 94.5 %

Serum 0.85 166 189 87.8 % 83.2 %

0.51 140 189 74.1 % 68.3 %

0.17 52 189 27.5 % 22.2 %

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Genotype verification
The performance of cobas® MPX to detect subtypes of HIV-1 Group M (A-H, J, K, BF, BG) and circulating recombinant
forms (CRF01_AE and CRF02_AG), HIV-1 Group O, HIV-1 Group N, and the subtypes of HIV-2 (A and B), genotypes of
HCV (1 - 6) and genotypes of HBV (A-H and precore mutant) was determined by testing unique clinical samples and/or
culture isolated for each subtype or genotype listed in Table 17 to Table 21.

HIV-1 Group M
A total of 115 unique HIV-1 Group M clinical samples with known HIV-1 subtype were quantified for HIV-1
concentrations using the COBAS® AmpliPrep/COBAS® TaqMan® HIV-1 Test, v2.0. All 115 samples were tested after
dilution with normal, virus-negative (HIV, HCV and HBV) human EDTA-plasma to 5 x LoD of cobas® MPX of which
102 samples were also tested neat (undiluted). All 115 clinical samples with known subtypes were detected neat and/or at
5 x LoD (Table 17).

Table 17 HIV-1 Group M clinical samples

Subtype % Reactive % Reactive

(reactive/samples tested) (reactive/samples tested)
neat diluted to 5 x LoD
A 100.0% (12/12) 100.0% (12/12)
CRF01_ AE 100.0% (12/12) 100.0% (12/12)
CRF02_AG 100.0% (12/12) 100.0% (12/12)
B 100.0% (11/11) 100.0% (11/11)
C 100.0% (12/12) 100.0% (12/12)
D 100.0% (11/11) 100.0% (11/11)
F 100.0% (10/10) 100.0% (10/10)
G 100.0% (12/12) 100.0% (12/12)
H 100.0% (10/10) 100.0% (10/10)
BF Not tested* 100% (3/3)
BG Not tested* 100% (4/4)
J Not tested* 100% (2/2)
K Not tested* 100% (4/4)

*Insufficient volume to test at neat

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HIV-1 Group O and HIV-1 Group N

A total of seven HIV-1 Group O and 2 HIV-1 Group N cultured isolates were tested after log dilutions were prepared in
normal, virus-negative (HIV, HCV and HBV) human EDTA-plasma. For HIV-1 Group O isolates, 28 total replicates
across seven isolates were tested using 4 replicates of each dilution. For HIV-1 Group N isolates, two isolates were tested. A
total of four replicates were tested for one isolate from dilution 1:1.00E+02 to 1:1.00E+03 and one replicate was tested for
the second isolate at dilution of 1:1.00E+04. HIV-1 Group O culture isolates were detected up to dilution of 1:1.00E+07 and
Group N culture isolates were detected up to dilution of 1:1.00E+04 (Table 18).

Table 18 HIV-1 Group O and HIV-1 Group N cultured isolates

% Reactive (reactive/valid replicates tested)

Sample Dilution
HIV-1 Group O HIV-1 Group N
1:1.00E+02 100.0% (28/28) 100.0% (4/4)
1:1.00E+03 100.0% (28/28) 100.0% (4/4)
1:1.00E+04 89.3% (25/28) 20.0% (1/5)
1:1.00E+05 71.4% (20/28) 0.0% (0/4)
1:1.00E+06 71.4% (20/28) 0.0% (0/4)
1:1.00E+07 71.4% (20/28) 0.0% (0/4)

HIV-2
A total of five HIV-2 subtype A (four) and B (one) cultured isolates were tested after log dilutions were prepared in
normal, virus-negative (HIV, HCV and HBV) human EDTA-plasma. For subtype A, a total of 16 replicates across four
isolates were tested for each dilution. For one isolate of subtype B, four total replicates were tested for each dilution. A total
of 11 HIV-2 subtype A (five) and B (six) clinical samples were also tested after log dilutions were prepared in normal,
virus-negative human EDTA-plasma. For subtype A, 20 total replicates across five clinical samples and for subtype B,
24 total replicates across six clinical samples were tested using four replicates for each dilution. All cultured isolates were
detected by cobas® MPX. Clinical samples were detected by cobas® MPX at up to dilutions of 1:1.00E+03 for subtypes A
and B. The overall results are summarized in Table 19.

Table 19 HIV-2 cultured isolates and clinical samples

% Reactive (reactive/valid replicates tested)

Sample Dilution Cultured isolate Clinical sample
Subtype A Subtype B Subtype A Subtype B
1:1.00E+02 100.0% (16/16) 100.0% (4/4) 100.0% (20/20) 100.0% (24/24)
1:1.00E+03 100.0% (16/16) 100.0% (4/4) 65.0% (13/20) 50.0% (12/24)
1:1.00E+04 100.0% (15/15) 100.0% (4/4) 25.0% (5/20) 0.0% (0/24)
1:1.00E+05 100.0% (16/16) 100.0% (4/4) 5.0% (1/20) 0.0% (0/24)
1:1.00E+06 100.0% (16/16) 100.0% (4/4) 0.0% (0/20) 0.0% (0/24)
1:1.00E+07 81.2% (13/16) 0.0% (0/4) 0.0% (0/20) 0.0% (0/24)

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HCV
A total of 96 unique HCV clinical samples with known HCV genotype were quantified for HCV concentrations using the
COBAS® AmpliPrep/COBAS® TaqMan® HCV Test, v2.0. All 96 HCV clinical samples with known genotypes were tested
after dilution with normal, virus-negative (HIV, HCV and HBV) human EDTA-plasma to 5 x LoD of cobas® MPX. Of
those, 95 samples were also tested neat. All samples were tested in single replicate. All 96 HCV-positive clinical samples
were detected neat and/or diluted as summarized in Table 20.
Table 20 HCV clinical samples

Genotype % Reactive % Reactive

(reactive/samples tested) (reactive/samples tested)
neat diluted to 5 x LoD
1a 100.0% (9/9) 100.0% (9/9)
1b 100.0% (12/12) 100.0% (12/12)
1 100.0% (12/12) 100.0% (12/12)
2b 100.0% (1/1) 100.0% (1/1)
2 100.0% (13/13) 100.0% (13/13)
3a 100.0% (12/12) 100.0% (12/12)
3 100.0% (1/1) 100.0% (1/1)
4 100.0% (13/13) 100.0% (13/13)
5a 100.0% (10/10) 100.0% (10/10)
5 100.0% (2/2) 100.0% (2/2)
6 100.0% (10/10) 100.0% (11/11)

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HBV
A total of 94 unique HBV clinical samples with known HBV genotype and pre-core mutants were quantified for HBV
concentrations using the COBAS® AmpliPrep/COBAS® TaqMan® HBV Test. All 94 HBV clinical samples with known
genotypes were tested neat and/or diluted with normal, virus-negative (HIV, HCV and HBV) EDTA-plasma to 5 x LoD of
cobas® MPX. All samples were tested with single replicates. All 94 HBV-positive clinical samples were detected both at
neat and/or diluted as summarized in Table 21.
Table 21 HBV clinical samples

Genotype % Reactive % Reactive

(reactive/samples tested) (reactive/ samples tested)
neat diluted to 5 x LoD
A 100.0% (15/15) 100.0% (15/15)
B 100.0% (12/12) 100.0% (11/11)
C 100.0% (10/10) 100.0% (9/9)
D 100.0% (12/12) 100.0% (11/11)
E 100.0% (12/12) 100.0% (11/11)
F 100.0% (12/12) 100.0% (12/12)
G Not tested* 100.0% (1/1)
H 100.0% (8/8) 100.0% (8/8)
Pre core Mutant 100.0% (12/12) 100.0% (12/12)

*Insufficient volume to test at neat

Seroconversion panels
The performance of cobas® MPX was evaluated using commercially available seroconversion panels for HIV-1 Group
M, HCV, and HBV. The results of cobas® MPX were compared to results for the same panels tested using the
cobas® TaqScreen MPX Test on the cobas s 201 system. In addition, a comparison was performed between cobas® MPX
and serology tests for each target.

HIV-1 Group M seroconversion panels

Twenty commercially available seroconversion panels were used. Each panel member was tested neat and diluted 1:6 and 1:96
to simulate testing in pools with cobas® MPX and cobas® TaqScreen MPX Test. The cobas® MPX results were compared to
the results obtained with the cobas® TaqScreen MPX Test and with results with the Abbott Architect HIV Ag/Ab Combo and
the Murex HIVAg/Ab Combo serology tests tested neat. The overall performance results are shown in Table 22.

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®
Table 22 Performance of cobas MPX on HIV seroconversion panels

Days earlier detection than HIV Antibody/Antigen or HIV RNA

®
Abbott ARCHITECT HIV Murex HIV Ab/Ag Combo: cobas TaqScreen MPX
HIV Seroconversion Ag/Ab Combo: Neat Neat Test: Neat, 1:6, 1:96
panels ®
Days earlier detection by the cobas MPX
Neat 1:6 1:96 Neat 1:6 1:96 Neat 1:6 1:96
1* 7 7 7 0 0 0 -4 0 -4
2 7 7 3 7 7 3 0 0 0
3 3 3 3 3 3 3 0 0 0
4 8 4 4 8 4 4 4 0 4
5 15 8 2 15 8 2 13 6 2
6 7 2 2 7 2 2 5 0 0
7 7 5 5 7 5 5 2 0 0
8 15 15 8 15 15 8 0 0 0
9 12 7 7 12 7 7 5 0 0
10 2 2 0 2 2 0 0 0 -2
11 6 0 0 6 0 0 6 0 0
12* 8 8 6 8 8 6 -5 2 0
13 7 7 7 7 7 7 0 0 2
14 10 3 3 10 3 3 2 0 0
15 12 7 7 12 7 7 0 -5 0
16 9 9 7 9 9 7 0 0 0
17 11 11 9 11 11 9 0 0 0
18 2 2 2 2 2 2 0 0 0
19 7 7 7 7 7 7 0 0 2
20 7 7 5 7 7 5 0 2 0
Minimum 2 0 0 0 0 0 -5 -5 -4
Average 8.1 6.1 4.7 7.8 5.7 4.4 1.4 0.3 0.2
Maximum 15 15 9 15 15 9 13 6 4

* Earlier reactivity was observed with the cobas® TaqScreen MPX Test due to HBV co-infection.

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HCV seroconversion panels

Twenty-five seroconversion panels were used. Each panel member was tested neat and diluted 1:6 and 1:96 to simulate
testing in pools with the cobas® MPX and cobas® TaqScreen MPX tests. The cobas® MPX results were compared to the
results obtained with the cobas® TaqScreen MPX Test and with results with the ORTHO HCV Version 3.0 ELISA Test
System and the Abbott PRISM HCV serology tests tested neat. The overall performance results are shown in Table 23.
®
Table 23 Performance of cobas MPX on HCV seroconversion panels

Days earlier detection than HCV Antibody/Antigen or HCV RNA

ORTHO HCV Version 3.0 cobas® TaqScreen MPX Test:
Abbott PRISM HCV: Neat
ELISA Test System: Neat Neat, 1:6, 1:96
Days earlier detection by the cobas® MPX

HCV Seroconversion panels Neat 1:6 1:96 Neat 1:6 1:96 Neat 1:6 1:96
1 13 13 13 7 7 7 0 0 0
2* 20 20 20 20 20 20 0 0 0
3 23 23 23 23 23 23 0 0 0
4* 23 23 23 19 19 19 0 0 0
5 33 33 33 33 33 33 -6 0 0
6* 39 39 39 37 37 37 0 0 0
7 32 32 32 32 32 32 0 0 0
8 38 38 38 38 38 38 -24** 0 0
9* 34 34 32 34 34 32 0 0 0
10* 32 32 29 32 32 29 0 3 0
11 34 34 34 34 34 34 0 0 0
12* 11 11 11 11 11 11 0 0 0
13* 10 10 10 10 10 10 0 0 0
14* 12 12 -2 12 12 -2 0 0 1
15 65 65 65 65 65 65 0 0 0
16 3 3 3 3 3 3 0 0 0
17* 13 13 13 16 16 16 0 0 0
18* 21 21 21 21 21 21 0 0 0
19 34 4 34 34 4 34 0 0 30
20 75 75 75 75 75 75 0 0 0
21 46 42 42 49 45 45 4 0 7
22 35 35 35 35 35 35 0 0 0

23 38 38 25 38 38 25 0 6 0
24 39 39 35 39 39 35 0 7 3
25 2 2 2 0 0 0 0 0 0
Minimum 2 2 -2 0 0 -2 -24 0 0
Average with exclusions* 34.0 31.7 30.4 33.7 31.4 30.1 -1.7 0.9 2.7
Maximum 75 75 75 75 75 75 4 7 30
* Panels were reactive on the first draw when tested with cobas® MPX or did not show seroconversion. These panels were
excluded from the summary calculations for the minimum, average and maximum number of days earlier detection than HCV
antibody or RNA for each dilution. One of the panels was used in 1:96 summary calculations versus serology only (panel 14).
** 24 day interval between adjacent draws.

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HBV seroconversion panels

Twenty-one commercially available seroconversion panels were used. Each panel member was tested neat and diluted 1:6
and 1:96 to simulate testing in pools with the cobas® MPX and cobas® TaqScreen MPX tests. The cobas® MPX results were
compared to the results obtained with the cobas® TaqScreen MPX Test and with results with the Genetic Systems HBsAg
and the Abbott PRISM HBsAg serology tests tested neat. The overall performance results are shown in Table 24.
®
Table 24 Performance of cobas MPX on HBV seroconversion panels

Days earlier detection than HBsAg or HBV DNA

Genetic Systems HBsAg cobas® TaqScreen MPX Test:

Abbott PRISM HBsAg: Neat
HBV Seroconversion panels (3.0): Neat Neat, 1:6, 1:96

Days earlier detection by the cobas® MPX

Neat 1:6 1:96 Neat 1:6 1:96 Neat 1:6 1:96

1 29 12 0 29 12 0 17 0 0
2 19 11 7 15 7 3 0 -3 0
3** 9 9 2 -14 -14 -21 0 0 0
4 38 27 19 38 27 19 4 0 2
5 22 0 0 22 0 0 0 -13 0
6 24 24 0 24 24 0 -7 7 0
7 21 18 7 21 18 7 3 4 0
8 21 14 11 21 14 11 3 0 7
9 19 12 5 19 12 5 0 5 5
10* 12 12 7 19 19 14 0 0 0
11** 17 17 0 0 0 -17 0 0 0
12 28 28 7 28 28 7 0 0 7
13 18 18 7 18 18 7 -8 4 10
14 18 15 7 11 8 0 9 0 5
15** 30 28 14 0 -2 -16 2 12 0
16 17 17 6 17 17 6 0 2 6
17 29 33 18 29 33 18 -4 15 3
18 22 10 0 22 10 0 12 0 0
19 18 14 3 18 14 3 4 0 0
20 28 28 0 28 28 0 -5 14 0
21 22 20 5 17 15 0 2 7 0

Minimum 9 0 0 -14 -14 -21 -8 -13 0

Average 22.5 17.8 6.0 18.2 13.5 2.2 1.6 2.7 2.1

Maximum 38 33 19 38 33 19 17 15 10

* Panel was consistently reactive with cobas® MPX, beginning on the first bleed and was excluded from the neat and 1:6
summary calculations for the minimum, average and maximum number of days earlier detection than HBV antibody.
**Low concentrations of HBV DNA were present in diluted panel members which were detected later by cobas® MPX than by
serology; 1.7 IU/mL in Panel 3 at 1:96, 2.0 IU/mL in Panel 11 at 1:96, and 0.5 IU/mL in Panel 15 at 1:96.

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Analytical specificity
The analytical specificity of cobas® MPX was evaluated for cross-reactivity with 25 microorganisms at 106 particles, copies,
or PFU/mL, which included 18 viral isolates, six bacterial strains and one yeast isolate (Table 25). The microorganisms
were added to normal, virus-negative (HIV, HCV and HBV) human EDTA-plasma and tested with and without HIV-1
Group M, HCV, HBV (co-formulated), HIV-1 Group O and HIV-2 virus added to a concentration of approximately
3 x LoD of cobas® MPX for each virus. The tested microorganisms do not cross-react or interfere with cobas® MPX.
Table 25 Microorganisms tested for analytical specificity

Viruses Flavivirus Bacteria Yeast

Adenovirus 5 West Nile Virus Escherichia coli Candida albicans
Cytomegalovirus Dengue Virus type 1 Propionibacterium acnes
Epstein-Barr Virus Usutu Virus Staphylococcus aureus
Herpes Simplex Virus type 1 Staphylococcus epidermidis
Herpes Simplex Virus type 2 Streptococcus viridans
Hepatitis A Virus Staphylococcus haemolyticus
Hepatitis E Virus
Hepatitis G Virus
Human T-cell lymphotropic Virus type I
Human T-cell lymphotropic Virus type II
Human Herpes Virus 6
Influenza Virus A
Parvovirus B19
Chikungunya Virus
Varicella Zoster Virus

Plasma samples from each of the disease states (Table 26) were tested with and without HIV-1 Group M, HCV, HBV
(co-formulated), HIV-1 Group O and HIV-2 added to a concentration of approximately 3 x LoD of cobas® MPX for
each virus. These disease states do not cross-react or interfere with cobas® MPX.

Table 26 Disease states samples tested for analytical specificity

Disease state
Adenovirus type 5 Herpes Simplex Virus type1 Human T-cell lymphotropic Virus type I
Cytomegalovirus Herpes Simplex Virus type 2 Human T-cell lymphotropic Virus type II
Dengue Virus Hepatitis A Virus Parvovirus B19
Epstein-Barr Virus Hepatitis E Virus West Nile Virus

Analytical specificity – interfering substances

Endogenous interference substances

Plasma samples with abnormally high levels of triglycerides (up to 33.2 g/L), hemoglobin (up to 2 g/L), unconjugated
bilirubin (up to 0.236 g/L), albumin (up to 60 g/L), and human DNA (up to 0.002 g/L) were tested with and without HIV-1
Group M, HCV, HBV (co-formulated), HIV-1 Group O and HIV-2 virus added to a concentration of 3 x LoD of cobas®
MPX. Samples containing these endogenous substances did not interfere with the sensitivity or specificity of cobas® MPX.

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Exogenous interference substances

Normal, virus-negative (HIV, HCV and HBV) human EDTA-plasma samples containing abnormally high concentrations
of drugs (Table 27) were tested with and without HIV-1 Group M, HCV, HBV (co-formulated), HIV-1 Group O and
HIV-2 added to a concentration of 3 x LoD of cobas® MPX for each virus. These exogenous substances did not interfere
with the sensitivity or specificity of cobas® MPX.

Table 27 Clinical samples tested with drugs

Name of drug tested Concentration

Acetaminophen 1324 µmol/L
Acetylsalicylic Acid 3620 µmol /L
Ascorbic Acid 342 µmol/L
Atorvastatin 600 µg Eq/L
Fluoxetine 11.2 µmol/L
Ibuprofen 2425 µmol/L
Loratadine 0.78 µmol/L
Nadolol 3.88 µmol/L
Naproxen 2170 µmol/L
Paroxetine 3.04 µmol/L
Phenylephrine HCL 491 µmol/L
Sertraline 1.96 µmol/L

Whole system failure

The whole system failure rate for cobas® MPX was determined by testing 100 replicates of EDTA plasma spiked with either
HIV-1 Group M, HCV, HBV (co-formulated), HIV-1 Group O, and HIV-2, for a total of 300 replicates. These samples
were tested at a target concentration of approximately 3 x LoD and were run in pools of 1 (undiluted). The study was
performed using the cobas® 8800 System with cobas p 680 instrument (pipetting and pooling).
The results of this study determined that all replicates were reactive for each target, resulting in a whole system failure rate of
0%. The two-sided 95% exact confidence interval was 0% for the lower bound and 1.22% for the upper bound [0%: 1.22%].

Cross contamination
The cross-contamination rate for cobas® MPX was determined by testing 240 replicates of a normal, virus-negative
(HIV, HCV and HBV) human EDTA-plasma sample and 220 replicates of a high titer HBV sample at 1.00E+08 IU/mL.
The study was performed using the cobas® 8800 System. In total, 5 runs were performed with positive and negative
samples in a checkerboard configuration.
All 240 replicates of the negative sample were non-reactive, resulting in a cross-contamination rate of 0%. The two-sided
95% exact confidence interval was 0% for the lower bound and 1.53% for the upper bound [0%: 1.53%].

FDA/CBER panel evaluation

The sensitivity of cobas® MPX was determined by testing FDA/CBER Lot Release panels for HIV-1 Group M, HIV-1
Group O, HIV-2, HBV and HCV. Panel members were tested undiluted in replicates across the reagent lots (one
replicate per panel member per reagent lot).

HIV-1 Group M
cobas® MPX is able to detect all levels of the FDA/CBER Lot Release Panel with concentrations of 10 cp/mL to
500 cp/mL and the negative panel member was non-reactive as summarized in Table 28.
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Table 28 Summary of FDA/CBER HIV-1 Group M RNA lot release panel results

Observed HIV Result

Concentration Expected
Panel Member
(cp/mL) Reactivity for HIV* Reagent Lot 1** Reagent Lot 2 Reagent Lot 3

B4 0 NR NT NR NR
B2 10 +/- NT R R
B6 50 +/- NT R R
B5 100 +/- NT R R
B9 500 R NT R R
* Reactivity: NR = non-reactive; +/- = may be reactive or non-reactive and were for information purposes only; R= reactive
** NT = Not Tested

HIV-1 Group O
cobas® MPX is able to detect all levels of the FDA/CBER Lot Release Panel with concentrations of 10 cp/mL to
1000 cp/mL as summarized in Table 29.

Table 29 Summary of FDA/CBER HIV-1 Group O RNA lot release panel results

Observed HIV Result

Concentration Expected
Panel Member
(cp/mL) Reactivity for HIV* Reagent Lot 1 Reagent Lot 2 Reagent Lot 3

Not
HIV-1 (O) #NC1 0 NR Not applicable** Not applicable**
applicable**
HIV-1 (O) #03 10 +/- R R R
HIV-1 (O) #02 100 +/- R R R
HIV-1 (O) #01 1000 R R R R

* Reactivity: NR = non-reactive; +/- = may be reactive or non-reactive and were for information purposes only; R= reactive
** Due to non-availability of panel member NC1 (negative panel member), this panel member could not be tested.

HIV-2
cobas® MPX is able to detect all levels of the FDA/CBER Lot Release Panel with concentrations of 5 cp/mL to
100 cp/mL and the negative panel member was non-reactive as summarized in Table 30.

Table 30 Summary of FDA/CBER HIV-2 RNA lot release panel results

Observed HIV Result

Concentration Expected
Panel Member
(cp/mL) Reactivity for HIV* Reagent Lot 1 Reagent Lot 2 Reagent Lot 3

HIV-2 #1 0 NR NR NR NR
HIV-2 #2 5 +/- R R R
HIV-2 #3 10 +/- R R R
HIV-2 #4 50 R R R R
HIV-2 #5 100 R R R R

* Reactivity: NR = non-reactive; +/- = may be reactive or non-reactive and were for information purposes only; R= reactive

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HBV
cobas® MPX is able to detect all levels of the FDA/CBER Lot Release Panel with concentrations of 10 cp/mL to
500 cp/mL and the negative panel member was non-reactive as summarized in Table 31.

Table 31 Summary of FDA/CBER HBV DNA lot release panel results

Observed HBV Result

Concentration Expected
Panel Member
(cp/mL) Reactivity for HBV* Reagent Lot 1 Reagent Lot 2 Reagent Lot 3

1 0 NR NR NR NR
2 10 +/- R R R
3 100 R R R R
4 50 +/- R R R
5 500 R R R R

* Reactivity: NR = non-reactive; +/- = may be reactive or non-reactive and were for information purposes only; R= reactive

HCV
cobas® MPX is able to detect all levels of the FDA/CBER Lot Release Panel with concentrations of 5 cp/mL to
500 cp/mL and the negative panel member was non-reactive as summarized in Table 32.

Table 32 Summary of FDA/CBER HCV RNA lot release panel results

Observed HCV Result

Concentration Expected
Panel Member
(cp/mL) Reactivity for HCV* Reagent Lot 1 Reagent Lot 2 Reagent Lot 3

2001 (#2) 0 NR NR NR NR
2002 (#10) 5 +/- R NR R
2003 (#9) 10 +/- R R NR
2004 (#8) 50 +/- R R R
2005 (#7) 100 R R R R
2006 (#6) 500 R R R R

* Reactivity: NR = non-reactive; +/- = may be reactive or non-reactive and were for information purposes only; R= reactive

The overall internal control failure rate across studies using living donor specimens was 0.46% with a 95% confidence
interval of 0.38% to 0.57%.

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Key performance characteristics - Cadaveric samples

Sensitivity
The clinical sensitivity of the cobas® MPX test for HIV-1 Group M RNA, HIV-1 Group O RNA, HIV-2, RNA HCV
RNA and HBV DNA was evaluated by testing a total of 60 individual virus-negative cadaveric samples, of those 35
individual samples were classified as moderately hemolyzed (straw to pink colored) and 25 individual samples were
classified as highly hemolyzed (red to brown colored). In addition a total of 60 individual virus-negative living donor
samples were tested. All cadaveric and living donor samples were divided evenly across 3 reagent lots, 5 clinical samples
spiking groups (for HIV-1 M, HCV and HBV) with 12 samples per group. Each cadaveric and living donor sample was
spiked with a co-formulation of three unique clinical samples (HIV-1 Group M, HCV and HBV), or Roche Primary
Standards (individually formulated HIV-1 Group O and HIV-2) at approximately 5 x LoD of respective sample types.
Each cadaveric sample was diluted 1:5.6 with cobas omni Specimen Diluent on the instrument and tested using the
cadaveric sample testing procedure.
All of the cadaveric and the living-donor samples had a reactive rate of 100% (95% confidence interval: 94.0 – 100%).
The clinical sensitivity observed in cadaveric sample was equivalent to the sensitivity observed in living donor samples
as determined by Fisher’s Exact Test and summarized in Table 33.

Table 33 Summary of reactivity rate in cadaveric and living donor samples in EDTA plasma

Cadaveric samples Living donor samples

Analyte % Reactive % Reactive

(Number of reactive /Number of samples (Number of reactive/Number of samples
tested) tested)

HIV-1 Group M 100% (60/60) 100% (60/60)

HIV-1 Group O 100% (60/60) 100% (60/60)
HIV-2 100% (60/60) 100% (60/60)
HCV 100% (60/60) 100% (60/60)
HBV 100% (60/60) 100% (60/60)
Fisher’s Exact Test, p-value
(α=0.05) No significant differences in reactive rates (p=1.000)

Specificity
The specificity of the cobas® MPX test in cadaveric EDTA plasma and serum samples was evaluated and compared with
the specificity in living donor sample by testing single replicates of 60 individual cadaveric EDTA plasma samples, of those
37 individual donor samples were classified as moderately hemolyzed (straw to pink colored) and 23 individual samples
were classified as highly hemolyzed (red to brown colored), 61 individual cadaveric serum samples of those 42 individual
samples were classified as moderately hemolyzed and 19 individual donor samples were classified as highly hemolyzed,
60 individual sero-negative living-donor plasma and 60 individual serum samples. The study was performed with
3 independent cobas® MPX reagent lots. Each cadaveric sample was diluted 1:5.6 with cobas omni Specimen Diluent on
the instrument and tested using the cadaveric sample testing procedure. All the cadaveric and living donor samples from
EDTA plasma and serum were non-reactive for 100% specificity. The specificity observed for cadaveric samples was equal
to the specificity observed for living-donor samples as determined by the Fisher’s Exact Test (α=0.05) as summarized in
Table 34.

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Table 34 Summary of specificity in cadaveric and living donor samples in EDTA plasma and serum

Matrices Sample type Number of non- Number of samples Two-sided 95%

% Non-reactive Confidence Interval
reactive tested

Cadaveric donor 60 94.0% - 100%

60 100%
EDTA plasma
Living donor 60 94.0% - 100%
60 100%

Cadaveric donor 94.1% - 100%

61 61 100%
Serum
Living donor 60 94.0% - 100%
60 100%

Overall results using Fisher’s Exact Test Specificity for cadaveric sample and living-donor samples are equivalent:
(α=0.05) Fisher’s Exact Test, p = 1.000

Reproducibility
The reproducibility of the cobas® MPX test on the cobas® 6800/8800 Systems was determined using 20 cadaveric samples
(moderately and highly hemolyzed) spiked with HIV-1 M, HBV and HCV clinical samples, and Roche Primary Standards
for HIV-1 Group O RNA and HIV-2 RNA to approximately 5 x LoD of the cobas® MPX test. The results were compared to
the reproducibility obtained with 20 living donor samples spiked with the Roche Primary and Secondary Standards to
approximately 5 x LoD of the cobas® MPX test.
Testing was performed for the following variable components:
 day-to-day variability over 6 days
 lot-to-lot variability using 3 different reagent lots of the cobas® MPX test
One replicate was tested with each of the 3 reagent lots over 6 days for a total of 18 replicates per cadaveric and living donor
sample. Each cadaveric sample was diluted 1:5.6 with cobas omni Specimen Diluent on the instrument and tested using
the cadaveric sample testing procedure. All valid reproducibility data were evaluated by comparing the reactive rates of
living donors and cadaveric samples (two-sided 95% Confidence Intervals) across all variable components. The Fisher’s
exact p value was calculated for the test of statistical significance of the difference between proportions of reactives
observed with cadaveric and living donor samples. No significant differences were observed.
cobas® MPX test is reproducible over multiple days and reagent lots for cadaveric and living donor samples. The results
from reagent lot-to-lot variability are summarized in Table 35.

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®
Table 35 cobas MPX test reagent lot-to-lot reproducibility summary for cadaveric and living donor samples

Significant
difference
% Reactive Lower limit Upper limit using Fisher’s
Reagent
Analyte Sample type (reactive/valid of 95% of 95% Exact Test
lot
replicates) confidence confidence
interval interval p-value
(α=0.05)
Cadaveric 100.0% (120/120) 97.0% 100.0%
1 p =1.0000
Living donor 100.0% (120/120) 97.0% 100.0%
Cadaveric 100.0% (120/120) 97.0% 100.0%
HIV-1 Group M 2 p =1.0000
Living donor 100.0% (120/120) 97.0% 100.0%
Cadaveric 100.0% (118/118) 96.9% 100.0%
3 p =1.0000
Living donor 100.0% (120/120) 97.0% 100.0%
Cadaveric 100.0% (120/120) 97.0% 100.0%
1 p =1.0000
Living donor 100.0% (120/120) 97.0% 100.0%
Cadaveric 100.0% (117/117) 96.9% 100.0%
HIV-1 Group O 2 p =1.0000
Living donor 100.0% (120/120) 97.0% 100.0%
Cadaveric 99.2% (118/119) 95.4% 100.0%
3 p =0.4979
Living donor 100.0% (120/120) 97.0% 100.0%
Cadaveric 100.0% (120/120) 97.0% 100.0%
1 p =1.0000
Living donor 100.0% (120/120) 97.0% 100.0%
Cadaveric 98.3% (118/120) 94.1% 99.8%
HIV-2 2 p =0.4979
Living donor 100.0% (120/120) 97.0% 100.0%
Cadaveric 99.2% (118/119) 95.4% 100.0%
3 p =0.4979
Living donor 100.0% (120/120) 97.0% 100.0%
Cadaveric 98.3% (118/120) 94.1% 99.8%
1 p =0.4979
Living donor 100.0% (120/120) 97.0% 100.0%
Cadaveric 98.3% (118/120) 94.1% 99.8%
HCV 2 p =0.4979
Living donor 100.0% (120/120) 97.0% 100.0%
Cadaveric 97.5% (115/118) 92.7% 99.5%
3 p =0.1203
Living donor 100.0% (120/120) 97.0% 100.0%
Cadaveric 100.0% (120/120) 97.0% 100.0%
1 p =1.0000
Living donor 100.0% (120/120) 97.0% 100.0%
Cadaveric 100.0% (120/120) 97.0% 100.0%
HBV 2 p =1.0000
Living donor 100.0% (120/120) 97.0% 100.0%
Cadaveric 100.0% (118/118) 96.9% 100.0%
3 p =1.0000
Living donor 99.2% (119/120) 95.4% 100.0%

The overall internal control failure rate across studies using cadaveric specimens was 3.39% with a 95% confidence
interval of 2.51% to 4.47%.
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Clinical performance evaluation

Reproducibility
The reproducibility of cobas® MPX for use on the cobas® 6800/8800 Systems was established by testing a 32-member
panel composed of two negative plasma samples and two plasma samples positive for each virus (HIV-1 Group M,
Group O, HIV-2, HCV, and HBV) at three different concentrations (approximately 0.5 x, 1.0 x, and 3.0 x the LoD of
cobas® MPX) for each virus.
Operators at each of the three cobas® MPX sites performed five days of testing, using three lots of cobas® MPX reagents to
obtain two valid batches per day. Two replicates per concentration were tested to yield up to 180 tests per panel member
virus type at each of the three concentrations.
All valid batches and test results were analyzed by calculating the percentage of reactive test results for each panel member
and the percentage of non-reactive results for the negative control panel member (Table 36). This study demonstrated that
cobas® MPX for use on the cobas® 6800/8800 Systems shows reproducible performance across the variables assessed (lot,
site/instrument, day, batch, and within batch) and for the five analytes tested.

Table 36 Test results summarized by site, lot, day, and batch (positive panel members)

Site Lot Day Batch

Viral
Load % % % %
Viral Concen- Positive Positive Positive Positive
Target tration ID Results ID Results ID Results ID Results
HIV-1 0.5 x LoD 1 81.7% (49/60) 1 81.7% (49/60) 1 91.7% (33/36) 1 84.3% (75/89)
Group M
2 84.7% (50/59) 2 88.3% (53/60) 2 77.1% (27/35) 2 81.1% (73/90)
3 81.7% (49/60) 3 78.0% (46/59) 3 83.3% (30/36)
4 83.3% (30/36)
5 77.8% (28/36)
1.0 x LoD 1 100.0% (60/60) 1 100.0% (60/60) 1 97.2% (35/36) 1 100.0% (90/90)
2 100.0% (60/60) 2 100.0% (60/60) 2 97.2% (35/36) 2 97.8% (88/90)
3 96.7% (58/60) 3 96.7% (58/60) 3 100.0% (36/36)
4 100.0% (36/36)
5 100.0% (36/36)
3.0 x LoD 1 100.0% (60/60) 1 100.0% (60/60) 1 100.0% (36/36) 1 100.0% (90/90)
2 100.0% (60/60) 2 100.0% (60/60) 2 100.0% (36/36) 2 100.0% (90/90)
3 100.0% (60/60) 3 100.0% (60/60) 3 100.0% (36/36)
4 100.0% (36/36)
5 100.0% (36/36)

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Site Lot Day Batch

Viral
Load % % % %
Viral Concen- Positive Positive Positive Positive
Target tration ID Results ID Results ID Results ID Results
HIV-1 0.5 x LoD 1 78.3% (47/60) 1 83.3% (50/60) 1 72.2% (26/36) 1 73.3% (66/90)
Group O
2 76.7% (46/60) 2 78.3% (47/60) 2 77.8% (28/36) 2 86.7% (78/90)
3 85.0% (51/60) 3 78.3% (47/60) 3 77.8% (28/36)
4 86.1% (31/36)
5 86.1% (31/36)
1.0 x LoD 1 98.3% (59/60) 1 98.3% (59/60) 1 94.4% (34/36) 1 95.6% (86/90)
2 100.0% (60/60) 2 96.7% (58/60) 2 100.0% (36/36) 2 98.9% (89/90)
3 93.3% (56/60) 3 96.7% (58/60) 3 97.2% (35/36)
4 100.0% (36/36)
5 94.4% (34/36)
3.0 x LoD 1 100.0% (60/60) 1 100.0% (60/60) 1 100.0% (36/36) 1 100.0% (90/90)
2 100.0% (60/60) 2 100.0% (60/60) 2 100.0% (36/36) 2 100.0% (90/90)
3 100.0% (60/60) 3 100.0% (60/60) 3 100.0% (36/36)
4 100.0% (36/36)
5 100.0% (36/36)
HIV-2 0.5 x LoD 1 74.1% (43/58) 1 73.3% (44/60) 1 77.8% (28/36) 1 69.7% (62/89)
2 76.7% (46/60) 2 79.7% (47/59) 2 69.4% (25/36) 2 79.8% (71/89)
3 73.3% (44/60) 3 71.2% (42/59) 3 75.0% (27/36)
4 71.4% (25/35)
5 80.0% (28/35)
1.0 x LoD 1 94.9% (56/59) 1 96.6% (57/59) 1 97.2% (35/36) 1 96.6% (85/88)
2 100.0% (59/59) 2 98.3% (58/59) 2 94.3% (33/35) 2 96.7% (87/90)
3 95.0% (57/60) 3 95.0% (57/60) 3 97.2% (35/36)
4 94.3% (33/35)
5 100.0% (36/36)
3.0 x LoD 1 100.0% (60/60) 1 100.0% (60/60) 1 100.0% (36/36) 1 100.0% (90/90)
2 100.0% (60/60) 2 100.0% (60/60) 2 100.0% (36/36) 2 100.0% (90/90)
3 100.0% (60/60) 3 100.0% (60/60) 3 100.0% (36/36)
4 100.0% (36/36)
5 100.0% (36/36)

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Site Lot Day Batch

Viral
Load % % % %
Viral Concen- Positive Positive Positive Positive
Target tration ID Results ID Results ID Results ID Results
HCV 0.5 x LoD 1 75.0% (45/60) 1 80.0% (48/60) 1 66.7% (24/36) 1 79.8% (71/89)
2 70.7% (41/58) 2 76.7% (46/60) 2 77.8% (28/36) 2 74.2% (66/89)
3 85.0% (51/60) 3 74.1% (43/58) 3 69.4% (25/36)
4 91.2% (31/34)
5 80.6% (29/36)
0.7  LoD 1 81.7% (49/60) 1 93.3% (56/60) 1 86.1% (31/36) 1 91.1% (82/90)
2 90.0% (54/60) 2 90.0% (54/60) 2 91.7% (33/36) 2 87.8% (79/90)
3 96.7% (58/60) 3 85.0% (51/60) 3 91.7% (33/36)
4 86.1% (31/36)
5 91.7% (33/36)
3.0  LoD 1 100.0% (60/60) 1 100.0% (60/60) 1 100.0% (36/36) 1 100.0% (90/90)
2 100.0% (59/59) 2 100.0% (60/60) 2 100.0% (36/36) 2 100.0% (89/89)
3 100.0% (60/60) 3 100.0% (59/59) 3 100.0% (36/36)
4 100.0% (35/35)
5 100.0% (36/36)
HBV 0.5  LoD 1 80.0% (48/60) 1 80.0% (48/60) 1 80.6% (29/36) 1 72.2% (65/90)
2 78.3% (47/60) 2 73.3% (44/60) 2 80.6% (29/36) 2 82.2% (74/90)
3 73.3% (44/60) 3 78.3% (47/60) 3 75.0% (27/36)
4 77.8% (28/36)
5 72.2% (26/36)
1.0  LoD 1 88.3% (53/60) 1 98.3% (59/60) 1 91.7% (33/36) 1 92.2% (83/90)
2 95.0% (57/60) 2 90.0% (54/60) 2 97.2% (35/36) 2 93.3% (84/90)
3 95.0% (57/60) 3 90.0% (54/60) 3 97.2% (35/36)
4 91.7% (33/36)
5 86.1% (31/36)
3.0  LoD 1 100.0% (60/60) 1 100.0% (60/60) 1 100.0% (36/36) 1 100.0% (90/90)
2 100.0% (60/60) 2 100.0% (60/60) 2 100.0% (36/36) 2 100.0% (90/90)
3 100.0% (60/60) 3 100.0% (60/60) 3 100.0% (36/36)
4 100.0% (36/36)
5 100.0% (36/36)

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Clinical specificity
Reactivity in blood donor population
Samples were collected from consented blood donors recruited from four test sites. Testing with cobas® MPX was done
according to two testing algorithms: one for individual donation testing, which required a single level of testing; and
one for pools of six testing, which required a single level of testing for primary pools that were non-reactive and
two levels of testing (primary pool and individual donation resolution testing for primary pools that were reactive). A
total of 74,180 donations were tested with cobas® MPX, resulting in an overall clinical specificity of 99.992%
(74,174/74,180; 99.982% to 99.996%) (Table 37). The pool specificity was 99.905% (10,524/10,534; 99.825%-99.948%)
(Table 38). Ten reactive pools contained all status negative donations. The clinical specificity for individual donation
testing was 99.946% (11,192/11,198; 99.883% to 99.975%). The invalid batch rate for the cobas® MPX was 3.5% for initial
testing donations in pools of six and for individual donations was 6.8% (16/235). Two HCV-positive NAT yield cases were
identified during this study.
®
Table 37 Clinical specificity of cobas MPX – overall

Estimate in Percent (95%

Pool Size Frequency (n/N) Score Confidence Interval)
Individual (Plasma) 5,523 / 5,528 99.910% (99.788% to 99.961%)
Individual (Serum) 5,669 / 5,670 99.982% (99.900% to 99.997%)
Individual (Plasma/Serum) 11,192 / 11,198 99.946% (99.883% to 99.975%)
Pools of 6 (Plasma) 62,982 / 62,982 100.000% (99.994% to 100.000%)
Overall 74,174 / 74,180 99.992% (99.982% to 99.996%)
®
N = Total number of status negative donations; n = cobas MPX non-reactive donations

®
Table 38 Pool reactivity of cobas MPX in volunteer blood donors

Percentage of
Category No. of Pools Pools Tested
Pools Tested 10,563 100
Non-Reactive pools 10,524 99.63
Reactive pools 39 0.37
Reactive pools with donor status positive 29 0.27
Reactive pools with donor status negative (false positive)* 10 0.10

* Of the 10 false reactive pools, one pool was HIV false reactive, four pools were HCV false reactive, and five pools were
HBV false reactive.

Reactivity in source plasma donor population

A total of 108,306 evaluable donations from 24,514 unique donors were tested in pools of 96 with both cobas® MPX
and cobas® TaqScreen MPX (incorporating the COBAS® AmpliScreen (CAS) tests for HBV, HCV, and HIV for viral
target resolution). One hundred eight thousand two hundred ninety-seven donations tested negative for anti-HIV,
anti-HCV, and HBsAg (Table 39). Donation status was assigned based on the concordance of two virus-specific tests
(e.g., two NAT results or NAT and serology) on the index donation or the results of follow-up testing. A total of
1,106 evaluable pools were tested with cobas® MPX, of which 1,092 (98.7%) were non-reactive and 14 (1.3%) were
reactive. Of the 1,092 non-reactive pools, 1,090 pools contained all status-negative donations, and two pools contained
at least one donation status-positive donation. Of the 1,106 pools tested, there were two non-reactive pools with at least
one status-positive donation and seven reactive pools with at least one status-positive donation (Table 40).

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®
Table 39 Clinical specificity of the cobas MPX – donation level
®
Total cobas MPX Result
Number of
Status-
Negative Estimate in Percent
Parameter Donations Reactive Non-Reactive (95% Score CI)

Clinical Specificity 108,297 6 108,291 99.994 (99.988, 99.997)

HIV Clinical Specificity 108,297 3 108,294 99.997 (99.992, 99.999)
HCV Clinical Specificity 108,297 1 108,296 99.999 (99.995, 100.000)
HBV Clinical Specificity 108,297 2 108,295 99.998 (99.993, 99.999)

Table 40 Pool reactivity in source plasma donations

Number Percentage of
Category of Pools Pools Tested
a
Total Pools of 96 tested: 1,106 100
b
Non-Reactive pools 1,092 98.7
Non-reactive pools with all donations status negative 1,090 98.6(1,090/1,106)
c
Non-reactive pools with at least one status-positive donation 2 0.2 (2/1,106)
b
Reactive pools 14 1.3
Reactive pools with at least one status-positive donation 7 0.6 (7/1,106)
Reactive pools with donation status-negative (false reactive pools) 7 0.6 (7/1,106)

a 479/1106 pools had < 96 donations

b Donation status was assigned based on the concordance of two virus-specific tests (e.g., two NAT results or NAT and
serology) on the index donation, or the results of follow-up testing.
c These two non-reactive pools contained donations from an HBV-positive donor. The donor’s index donation was HBV-
® ®
positive on cobas MPX but negative on cobas TaqScreen MPX Test and was confirmed HBV-positive by alternative
high-sensitivity NAT. This donor made three subsequent donations that were nonreactive on both NAT screening assays.
One of these donations was contained within an HCV-positive pool.
Eleven unique donors contributed 12 reactive donations (six HCV, six HIV, and three HBV). Seven donors completed
follow-up testing: three of these donors did not show evidence of infection on follow-up; four donors were confirmed to
have infection on follow-up, of whom two seroconverted (HCV) during follow-up (Table 41). One of the three HBV
donors was determined to be a NAT HBV yield case.

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Table 41 Observed testing reactivity patterns from initial testing on evaluable donations

Donation Number of
® a
cobas MPX Result Status Donations
HCV+ Positive 5
HBV+ Negative 2
b
HBV+ Positive 4
HCV+ Negative 1
HIV+ Negative 3
Non-Reactive Negative 108,291
Total 108,306

a Donation Status was assigned based on the test reactivity pattern (concordance” of two virus-specific tests (e.g., two
NAT results or NAT and serology) on the index donation or results of follow-up testing).
b These donations are all from the same donor whose index donation was HBV+ and whose subsequent three donations
were classified as status positive even though cobas MPX test was non-reactive for HBV.
Note: Only evaluable donations are included in this summary table; + = Reactive/Positive
The clinical specificity of cobas® MPX for source plasma pools was determined by the analysis of 108,306 evaluable donations
from 24,514 unique donors. Evaluable donations had valid cobas® MPX, cobas® TaqScreen MPX Test and CAS results from
testing pools, and valid serology results (across analytes) from testing of individual donations. Of these 108,306 evaluable
donations, 108,297 were assigned a donation status of negative, of which 108,291 were cobas® MPX non-reactive, for a
clinical specificity of 99.994% (95% Confidence Interval: 99.988% to 99.997%). Seven false cobas® MPX reactive pools of 96
resolved to contain all status-negative donations. Of the 24,514 unique donors tested, 24,509 contributed only status-negative
donations, of which 24,503 were non-reactive on cobas® MPX and six had false-reactive results, resulting in specificity (at the
donor level) of 99.976% (95% Confidence Interval: 99.947% to 99.989%).

Studies in high risk populations

Third-party vendors collected samples from individuals at high risk for infection with HIV, HCV, or HBV. High-risk
factors were included, but were not limited to, a history of incarceration; history of a diagnosis of a sexually-transmitted
disease; history of multiple sex partners; use of injection drugs; diagnosed with or treated for HIV; and diagnosed with or
treated for hepatitis. Some sample contributors indicated more than one risk factor. A total of 510 samples from a high
risk population were distributed approximately evenly across three test sites and tested with cobas® MPX and
cobas® TaqScreen MPX incorporating CAS.
All samples were prepared as panels. The diluted samples were manually diluted with pooled human plasma confirmed
to be negative for HIV-1/2, HCV, and HBV. At the testing sites, samples were tested neat with both cobas® MPX and
cobas® TaqScreen incorporating CAS (for target resolution), as per the Standard Specimen Processing Procedure
recommended in the cobas® TaqScreen MPX Test Package Insert. Samples were also tested dilute to simulate pools of
six with both cobas® MPX and cobas® TaqScreen. CAS was not performed on dilute samples.
The 510 neat samples generated results from cobas® MPX and the cobas® TaqScreen MPX Test which included
179 samples reactive (for one or more targets) on cobas® MPX (35.1%); and 181 samples that were reactive on
cobas® TaqScreen MPX Test (35.5%). 488 (95.7%) samples that showed results concordant between cobas® MPX and
cobas® TaqScreen MPX Test, while 22 (4.3%) of samples produced results that were discordant between cobas® MPX and
cobas® TaqScreen MPX Test.
For the 510 high-risk neat samples, cobas® MPX correctly identified the presence or absence of viral target 97.0%
(495/510) of the time, compared to CAS or alternative NAT (NGI; National Genetics Institute) test results. For the 3% of
samples for which cobas® MPX did not correctly identify the presence or absence of viral target, cobas® MPX incorrectly
detected a viral target in samples that did not contain a viral target 1.8% (9/510) of time (false reactive result) and failed to
detect a viral target in samples that contained a target 1.2% (6/510) of the time (false non-reactive result). These results are
summarized in Table 42.

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Table 42 Correct versus incorrect identification of virus - neat

® a
cobas MPX Result % Total Correct
True reactives 170
97.0% 495
True non-reactives 325
False reactives 9 1.8%
15
False non-reactives 6 1.2%
Total 510 100.0% 510

a Final status (as compared with CAS or alternative NAT [NGI testing] results)
Note: Correct identification = True reactive and true non-reactive results (shown in bold type).

Of 510 dilute samples tested, 153 samples were reactive on cobas® MPX (30.0%), compared to 151 samples that were
reactive on cobas® TaqScreen MPX Test (29.6%). Of the 510 dilute samples, 484 (94.9%) samples showed results
concordant between cobas® MPX and cobas® TaqScreen MPX Test; and 26 (5.1%) samples showed results discordant
between cobas® MPX and cobas® TaqScreen MPX Test.
cobas® MPX correctly identified the viral target 96.7% (492/509) of the time (509 dilute samples excludes one sample for
which no NGI result was obtained). For the 3.4% of samples for which cobas® MPX did not correctly identify the viral
target, cobas® MPX incorrectly detected a viral target in samples that did not contain a viral target 1.2% (6/509) of time
(false reactive result) and failed to detect a viral target in samples that contained a target 2.2% (11/509) of the time (false
non-reactive result). These results are summarized in Table 43.

Table 43 Correct versus incorrect identification of virus - dilute

® a
cobas MPX Result % Total correct
True reactives 147
96.7 492
True non-reactives 345
False reactives 6 1.2
17
False non-reactives 11 2.2
b b
Total 509 100 509

a Final status (as compared with CAS or alternative NAT [NGI testing] results), which was performed on neat aliquot.
b Excludes one sample for which no NGI result was obtained.
Note: Correct identification = True reactive and true non-reactive results (shown in bold type).

Clinical sensitivity

Studies in NAT-positive populations

A total of 2,569 HIV, HCV, and HBV NAT-positive samples were tested across four test sites with cobas® MPX and the
cobas® TaqScreen MPX Test incorporating CAS. Four lots of cobas® MPX reagents were used. The 2,569 samples known
to be NAT-positive consisted of 1,015 HIV-positive samples, 1,016 HCV-positive samples, and 538 HBV-positive samples.
Each of these samples were tested both neat and dilute (1:6) with cobas® MPX and the cobas® TaqScreen MPX Test. Only
neat, not dilute, samples, were tested with the licensed CAS Tests per the Standard Specimen Processing Procedure
recommended in the cobas® TaqScreen MPX Test Package Insert. Table 44 compares the sensitivities of cobas® MPX and
cobas® TaqScreen Test Results for HIV, HCV, and HBV Known Positive Samples.
The overall clinical sensitivity of the cobas® MPX was 100.0% (2,549/2,549) for neat known positive samples and 100.0%
(2,555/2,555) for dilute (1:6) known positive samples. The overall clinical sensitivity of the cobas® TaqScreen MPX Test
was 99.9% (2,523/2,524) for neat known positive samples and 99.8% (2,559/2,563) for dilute (1:6) known positive
samples. The overall positive percent agreement (PPA) across all known positive samples in this study between
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cobas® MPX and the cobas® TaqScreen MPX Test was 100.0% for both neat and dilute samples (Table 44).
® ®
Table 44 Comparison of the sensitivities of cobas MPX and cobas TaqScreen Test results for HIV, HCV, and HBV known positive samples

Difference
Sensitivity in ® ®
a (cobas MPX Result − cobas
Known Positive Samples
TaqScreen MPX Test)
®
Target ® cobas TaqScreen MPX 95% Confidence
Dilution cobas MPX Result Estimate
Virus Test Interval
Overall 100.00% (2,549/2,549) 99.96% (2,523/2,524) 0.04% (-0.04%, 0.12%)

Neat HIV 100.00% (1,006/1,006) 99.90% (1,007/1,008) 0.10% (-0.10%, 0.29%)

HCV 100.00% (1,015/1,015) 100.00% (1,014/1,014) 0.00% Not applicable
HBV 100.00% (528/528) 100.00% (502/502) 0.00% Not applicable
Overall 100.00% (2,555/2,555) 99.84% (2,559/2,563) 0.16% (0.00%, 0.31%)

1:6 HIV 100.00% (1,006/1,006) 99.60% (1,005/1,009) 0.40% (0.01%, 0.78%)

HCV 100.00% (1,016/1016) 100.00% (1,016/1,016) 0.00% Not applicable
HBV 100.00% (533/533) 100.00% (538/538) 0.00% Not applicable
a Only known positive samples with valid test results were included in the sensitivity analysis.

HIV NAT positive population

The 1,015 HIV-positive neat samples generated 1,006 evaluable test results with cobas® MPX and 1,008 evaluable test
results with the cobas® TaqScreen MPX Test incorporating CAS. One thousand fifteen HIV dilute samples produced
1,006 evaluable test results with cobas® MPX and 1,009 evaluable test results with the cobas® TaqScreen MPX Test (CAS
was not performed on dilute samples).
cobas® MPX was reactive for 1,006 of 1,006 (100.0%) HIV neat samples and 1,006 of 1,006 (100.0%) HIV dilute
samples. The cobas® TaqScreen MPX Test incorporating CAS was reactive for 1,007 of 1,008 (99.90 %) for HIV neat
samples. The cobas® TaqScreen MPX Test (no CAS performed) was reactive for 1,005 of 1,009 (99.60%) for HIV dilute
samples. The PPA between cobas® MPX and the cobas® TaqScreen MPX Test for neat and dilute HIV samples was
100.0% and 100.0% respectively.

HCV NAT positive population

cobas® MPX was reactive for 1,015 of 1,015 (100.0%) HCV neat samples and 1,016 of 1,016 (100.0%) HCV dilute
samples. The cobas® TaqScreen MPX Test incorporating CAS was also reactive for 1,014 of 1,014 (100.0 %) for neat
samples. The cobas® TaqScreen MPX Test (no CAS performed) was reactive for 1,016 of 1,016 (100.0%) for dilute
samples. The PPA between cobas® MPX and the cobas® TaqScreen MPX Test for neat and dilute HCV samples was
100.0% and 100.0% respectively.

HBV NAT-positive population

The 538 HBV-positive neat samples generated 528 evaluable test results with cobas® MPX and 502 evaluable test results with the
cobas® TaqScreen MPX Test incorporating CAS. The 538 HBV dilute samples produced 533 evaluable test results with
cobas® MPX, and 538 evaluable test results with the cobas® TaqScreen MPX Test (CAS was not performed on dilute samples).
cobas® MPX was reactive for 528 of 528 (100.0%) HBV-positive neat samples and 533 of 533 (100.0%) HBV-positive
dilute samples. The cobas® TaqScreen MPX Test incorporating CAS was reactive for 502 of 502 (100.0%) for HBV neat
samples. The cobas® TaqScreen MPX Test (no CAS performed) was reactive for 538 of 538 (100.0%) for HBV dilute
samples. The PPA between cobas® MPX and the cobas® TaqScreen MPX Test for neat and dilute HBV samples was
100.0% and 100.0% respectively.

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Clinical sensitivity for HIV-1 Group O and HIV-2 seropositive population

HIV-1 Group O seropositive population

A total of 12 HIV-1 Group O seropositive samples were tested after 1:6 dilution using cobas® MPX and cobas® TaqScreen
MPX Test. The samples were tested after 1:6 dilution due to limited volume. All of the HIV-1 Group O samples were
reactive for HIV when tested with cobas® MPX after a 1:6 dilution as summarized in Table 45, for a clinical sensitivity of
100.0% relative to serology.

Table 45 Comparison of overall reactivity for HIV-1 Group O seropositive samples (1:6 dilution)
®
®
cobas TaqScreen MPX Test cobas MPX (1:6 Dilution)
Total
(1:6 Dilution) Reactive Non-Reactive

Reactive 11 0 11

Non-Reactive 1 0 1

Total 12 0 12

HIV-2 seropositive population

A total of 319 HIV-2 seropositive samples were tested using the cobas® MPX and cobas® TaqScreen MPX Test. Out of the
319 seropositive samples, 184 were tested neat and after 1:6 dilution with cobas® MPX and cobas® TaqScreen MPX Test
whereas the remaining 135 were only tested after 1:6 dilution due to limited volume.
A total of 137 samples of the 184 neat tested samples was reactive as summarized in Table 46, for a clinical sensitivity of
74.5% relative to serology using cobas® MPX. Comparable sensitivity of cobas® MPX towards HIV-2 was also
demonstrated when samples were diluted 1:6 prior to testing with both methods. A total of 198 samples of the 319 1:6
diluted samples were reactive with cobas® MPX as summarized in Table 47.

Table 46 Comparison of overall reactivity for HIV-2 seropositive samples (neat)

®
®
cobas TaqScreen MPX Test cobas MPX (Neat)
Total
(Neat) Reactive Non-Reactive

Reactive 118 7 125

Non-Reactive 19 40 59

Total 137 47 184

Table 47 Comparison of overall reactivity for HIV-2 seropositive samples (1:6 dilution)
®
®
cobas TaqScreen MPX Test cobas MPX (1:6 Dilution)
Total
(1:6 Dilution) Reactive Non-Reactive

Reactive 173 33 206

Non-Reactive 25 88 113

Total 198 121 319

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Confirmation of serology results

Data from the Known Positive Study included 2,555 known-positive samples, each with nucleic acid test (NAT)-confirmed
infection with either HIV, HCV, or HBV and serology test results. Supplemental serology test results were also known for
1,771 (69.3%) samples. The correct cobas® MPX result, defined as reactive for the viral target for which the specimen was
known to be positive (e.g., HIV, HCV, or HBV), was compared to the supplemental serology results. The percentages of
correct results (sensitivity estimate) for cobas® MPX were calculated for each target virus and overall, with associated 95%
confidence intervals (CI). cobas® MPX correctly identified 1,771 of 1,771 (100.0%) of specimens with reactive serology
and supplemental serology results. Table 48 shows the reactivity of cobas® MPX for each viral target analyte, compared to
the known viral target serology and supplemental serology test result, as well as an estimate of sensitivity and 95% CI
overall and for each viral target.
®
Table 48 Sensitivity of the cobas MPX for neat known positive specimens with confirmatory serology results

Total Known Number

Target Positive Reactive Sensitivity
Dilution Test Virus Specimens* By Test Estimate 95% Score CI
Neat MPX8800 Overall 1,771 1,771 100.00% (99.78%, 100.00%)
Neat MPX8800 HIV 496 496 100.00% (99.23%, 100.00%)
Neat MPX8800 HCV 747 747 100.00% (99.49%, 100.00%)
Neat MPX8800 HBV 528 528 100.00% (99.28%, 100.00%)
®
* Only known positive specimens with valid cobas MPX results from neat samples and confirmatory serology results are
included in this sensitivity analysis.

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Additional information
Key test features
Sample type Plasma and Serum
Minimum amount of sample required for living donor 1000 µL
Amount of sample processed for living donor 850 µL
Minimum amount of sample required for cadaveric donor 300 µL
Amount of sample processed for cadaveric donor 150 µL
Results are available within less than 3.5 hours
Test duration
after loading the sample on the system.

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Symbols
The following symbols are used in labeling for Roche PCR diagnostic products.

Table 49 Symbols used in labeling for Roche PCR diagnostics products

Ancillary Software In Vitro Diagnostic Medical Device

Authorized Representative
Lower Limit of Assigned Range
in the European community

Barcode Data Sheet Manufacturer

Batch code Store in the dark

Biological Risks Contains sufficient for <n> tests

Catalogue number Temperature Limit

Consult instructions for use Test Definition File

Contents of kit Upper Limit of Assigned Range

Distributed by Use-by date

For IVD Performance Evaluation

Global Trade Item Number
Only

This product fulfills the requirements of the European Directive 98/79 EC for in vitro diagnostic
medical devices.

US Customer Technical Support 1-800-526-1247

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Manufacturer and distributors

Table 50 Manufacturer and distributors
Manufactured in Switzerland
Roche Diagnostics GmbH
Sandhofer Strasse 116
68305 Mannheim, Germany
www.roche.com

Roche Diagnostics
201, boulevard Armand-Frappier
H7V 4A2 Laval, Québec, Canada
(For Technical Assistance call:
Pour toute assistance technique,
appeler le: 1-877-273-3433)

Trademarks and patents

See http://www.roche-diagnostics.us/patents

Copyright
©2017 Roche Molecular Systems, Inc.

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Document revision
Document Revision Information
Doc Rev. 2.0 Addition of cadaveric claims.
11/2017 Updated Sample collection, transport and storage section to reflect extended frozen
storage condition.
Added Greiner Gel Tubes for sample collection, transport and storage.
Updated complete information of MPX Master Mix Reagent 2 in Table 1.
Updated the P/N for the cobas p 680 Instrument.
Added Roche web address www.roche.com.
Please contact your local Roche Representative if you have any questions.

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