OnSite Toxo IgG/IgM Combo Rapid Test - (Serum / Plasma / Whole Blood) Page 1 of 2

10. Dispose of all specimens and materials used to perform the test as bio-hazardous waste.
OnSite™ Toxo IgG/IgM Combo Rapid Test 11.
12.
Handle negative and positive controls in the same manner as patient specimens.
The test result should be read 10-15 minutes after a specimen is applied to the sample well or
sample pad of the device. Any results interpreted outside of the 10-15 minute window should
be considered invalid and must be repeated.
R0234C 13. Do not perform the test in a room with strong air flow, e.g. an electric fan or strong air
conditioning.
INTENDED USE REAGENT PREPARATION AND STORAGE INSTRUCTIONS
The OnSite Toxo IgG/IgM Combo Rapid Test is a lateral flow immunoassay for the simultaneous All reagents are ready to use as supplied. Store unused test devices unopened at 2-30°C. If stored at
detection and differentiation of IgM and IgG antibodies to Toxoplasma gondii (T. gondii) in human 2-8°C, ensure that the test device is brought to room temperature before opening. The test device is
serum, plasma or whole blood. It is intended to be used by professionals as a screening test and stable through the expiration date printed on the sealed pouch. Do not freeze the kit or expose the kit to
provides a preliminary test result to aid in the diagnosis of infection with T. gondii. temperatures above 30°C.
Any interpretation or use of this preliminary test result must also rely on other clinical findings as well as SPECIMEN COLLECTION AND HANDLING
on the professional judgment of health care providers. Alternative test method(s) should be considered
to confirm the test result obtained by this device. Consider any materials of human origin as infectious and handle them using standard bio-safety
procedures.
SUMMARY AND EXPLANATION OF THE TEST
Plasma

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T. gondii is an obligate intracellular protozoan parasite with a worldwide distribution1,2. Serological data
Step 1: Collect blood specimen into a lavender, blue or green top collection tube (containing EDTA,
indicates that approximately 6-47% of the population is chronically infected with the organism3. A T.
citrate or heparin, respectively, in Vacutainer®) by venipuncture.
gondii infection occurs essentially without knowledge of the patient and may be unrelated to direct
Step 2: Separate the plasma by centrifugation.
exposure to a cat (e.g., by ingestion of vegetables or water contaminated with oocysts or ingestion of
Step 3: Carefully withdraw the plasma into new pre-labeled tube.

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undercooked meat contaminated with cysts)4. An initial T. gondii infection and the subsequent chronic
infection are clinically undetected in 80% to 89% of healthy individual5. In immunosuppressed patients, Serum
such as patients infected with HIV, both acute and recurrent toxoplasmosis can have severe clinical Step 1: Collect blood specimen into a red top collection tube (containing no anticoagulants in
manifestations5. In pregnant women, an acute T. gondii infection may lead to serious fetal congenital Vacutainer®) by venipuncture.
mental retardation, blindness and hydrocephaly6. Step 2: Allow the blood to clot.

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Step 3: Separate the serum by centrifugation.
Methods for diagnosis of toxoplasmosis include PCR, histological diagnosis, parasite isolation and
Step 4: Carefully withdraw the serum into a new pre-labeled tube.
serology4. Serological detection of T. gondii specific IgM and IgG antibodies is the primary method for
diagnosis of toxoplasmosis. Diagnosis of toxoplasmosis is helpful in determining the risk for congenital Test specimens as soon as possible after collecting. Store specimens at 2-8°C, if not tested
toxoplasmosis during pregnancy4. immediately for up to 5 days. The specimens should be frozen at -20°C for longer storage.
IgM anti-T. gondii develops during acute primary infection with T. gondii and declines generally within a Avoid multiple freeze-thaw cycles. Prior to testing, bring frozen specimens to room temperature slowly
few months4. IgG anti-T. gondii develops generally within 1–2 weeks post infection, peaks within 1–2 and mix gently. Specimens containing visible particulate matter should be clarified by centrifugation
months, and usually persists for life4. The absence of IgG antibodies before or early in pregnancy before testing. Do not use specimens demonstrating gross lipemia, gross hemolysis or turbidity in order

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allows the identification of women at risk of acquiring infection7. Additionally, the presence of IgG allows to avoid interference with result interpretation.
the identification of immunocompromised patients at risk for the reactivation of a latent infection7.
Whole Blood
The OnSite Toxo IgG/IgM Combo Rapid Test detects IgM and IgG anti-T. gondii in human serum,
Drops of whole blood can be obtained by either finger tip puncture or venipuncture. Collect blood
plasma or whole blood by utilizing T. gondii-specific antigens. The test can be performed within 10
specimen into a lavender, blue or green top collection tube (containing EDTA, citrate or heparin,
minutes by minimally skilled personnel without the use of laboratory equipment.
respectively in Vacutainer®). Do not use hemolyzed blood for testing.
TEST PRINCIPLE Whole blood specimens should be stored at 2-8°C if not tested immediately. The specimens must be
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The OnSite Toxo IgG/IgM Combo Rapid Test is a tested within 24 hours of collection.
lateral flow chromatographic immunoassay. The test ASSAY PROCEDURE
strip in cassette device consists of: 1) a burgundy
colored conjugate pad containing a recombinant T. S Step 1: Bring the specimen and test components to room temperature if refrigerated or frozen. Once
gondii antigen conjugated with colloidal gold (Toxo the specimen is thawed, mix well prior to performing the assay.
conjugates) and a control antibody conjugated with
colloidal gold and 2) a nitrocellulose membrane strip Step 2: When ready to test, open the pouch at the notch and remove the device. Place the test device
on a clean, flat surface.
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containing two test lines (G and M lines) and a control line (C line). The G line is pre-coated with anti-
human IgG for detection of IgG anti-T. gondii. The M line is pre-coated with mouse anti-human IgM for
Step 3: Be sure to label the device with the specimen’s ID number.
detection of IgM anti-T. gondii. The C line is pre-coated with a control antibody.
Step 4: Fill the capillary tube with specimen not exceeding the specimen line as shown in the images
When an adequate volume of test specimen and sample diluent is dispensed into the sample well and
below. The volume of specimen is approximately 10 µL. For better precision, transfer
buffer well, respectively, the specimen migrates by capillary action across the test strip. IgM anti-T.
specimen using a pipette capable of delivering a 10 µL volume.
gondii, if present in the specimen, will bind to the Toxo conjugates. The immunocomplex is then
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captured on the membrane by the pre-coated anti-human IgM forming a burgundy colored M line, Holding the capillary tube vertically, dispense the entire specimen into the center of the
indicating an IgM anti-T. gondii positive test result. IgG anti-T. gondii, if present in the specimen, will sample well making sure that there are no air bubbles.
bind to the Toxo conjugates. The immunocomplex is then captured on the membrane by the pre-coated
anti-human IgG forming a burgundy colored G line, indicating an IgG anti-T. gondii positive test result. Immediately add 2 drops (about 60-80 µL) of sample diluent to the sample well with bottle
positioned vertically.
Absence of any test lines (G or M) suggests a negative test result. The test contains an internal control
(C line) which should exhibit a burgundy colored line of the immunocomplex of the control antibodies,
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regardless of color development on the test lines (G and M). If no control line (C line) develops, the test
result is invalid and the specimen must be retested with another device.

REAGENTS AND MATERIALS PROVIDED

1. Individually sealed foil pouches containing:

a. One cassette device 10 µL serum or plasma 2 drops of sample diluent
b. One desiccant
Result
2. 10 µL capillary tubes 10 minutes
3. Sample diluent (REF SB-R0234, 5 mL/bottle)
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4. One package insert (instructions for use)

MATERIALS MAY BE REQUIRED BUT NOT PROVIDED

1. Positive control
2. Negative control
10 µL whole blood 2 drops of sample diluent
MATERIALS REQUIRED BUT NOT PROVIDED
Step 5: Set up the timer.
1. Clock or timer
2. Lancing device for whole blood test Step 6: Result should be read at 10 minutes. Positive results may be visible in as soon as 1 minute.
Negative results must be confirmed at the end of 15 minutes only. Any results interpreted
WARNINGS AND PRECAUTIONS outside of the 10-15 minute window should be considered invalid and must be
repeated. Discard used devices after interpreting the result following local
For In Vitro Diagnostic Use
requirements governing the disposal of devices.
1. This package insert must be read completely before performing the test. Failure to follow the
insert may lead to inaccurate test results. QUALITY CONTROL
2. Do not open the sealed pouch until ready to conduct the assay.
3. Do not use expired devices or components. 1. Internal Control: This test contains a built-in control feature, the C line. The C line develops
4. Bring all reagents to room temperature (15-30°C) before use. after adding the specimen and the sample diluent. If the C line does not develop, review the
5. Do not use components from another test kit to substitute for components of this kit. entire procedure and repeat the test with a new device.
6. Do not use hemolyzed blood specimens for testing.
2. External Control: Good Laboratory Practice recommends using external controls, positive
7. Wear protective clothing and disposable gloves while handling the kit reagents and clinical
and negative, to assure the proper performance of the assay, particularly under the following
specimens. Wash hands thoroughly after performing the test.
circumstances:
8. Users of this test should follow the US CDC Universal Precautions for prevention of
a. A new operator uses the kit, prior to performing the testing of the specimens.
transmission of HIV, HBV and other blood-borne pathogens.
b. A new lot of test kits is used.
9. Do not smoke, drink or eat in areas where specimens or kit reagents are being handled.

Copyright 2016 by CTK Biotech, Inc.

 OnSite Toxo IgG/IgM Combo Rapid Test - (Serum / Plasma / Whole Blood) Page 2 of 2

c. A new shipment of test kits is used. 6. Interference

d. The temperature during storage of the kits falls outside of 2-30°C. Common substances (such as pain and fever medication and blood components) may affect the
e. The temperature of the test area falls outside of 15-30°C. performance of the OnSite Toxo IgG/IgM Combo Rapid Test. This was studied by spiking these
f. To verify a higher than expected frequency of positive or negative results. substances into IgM positive, medium-level IgG positive, weak-level IgG positive, and IgM and
g. To investigate the cause of repeated invalid results. IgG negative specimens, respectively. The results demonstrate that at the concentrations
tested, the substances studied do not affect the performance of the OnSite Toxo IgG/IgM
INTERPRETATION OF ASSAY RESULT
Combo Rapid Test.
1. NEGATIVE RESULT: If only the C line develops, the test indicates that anti-T. gondii antibodies
List of potentially interfering substances and concentrations tested:
are not detected in the specimen. The result is negative or non-reactive.
1. Albumin 60 g/L 6. Hemoglobin 2 g/L
2. Bilirubin 20 mg/dL 7. Heparin 3,000 U/L
3. Creatinine 442 µmol/L 8. Salicylic acid 4.24 mmol/L
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4. EDTA 3.4 µmol/L 9. Sodium citrate 3.8%
5. Glucose 55 mmol/L

2. POSITIVE RESULT: LIMITATIONS OF TEST

2.1 In addition to the presence of the C line, if only the M line is developed, the test indicates the
1. The Assay Procedure and the Interpretation of Assay Result sections must be followed closely
presence of IgM anti-T. gondii. The result is IgM anti-T. gondii positive or reactive and IgG anti-
when testing for the presence of antibodies to T. gondii in serum, plasma or whole blood from
T. gondii negative or non-reactive.
individual subjects. Failure to follow the procedure may lead to inaccurate test results.

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2. The OnSite Toxo IgG/IgM Combo Rapid Test is limited to the qualitative detection of antibodies
to T. gondii in serum, plasma or whole blood. The intensities of the test lines do not have linear
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correlation with the antibody titers in the specimen.
3. A negative or non-reactive result for an individual subject indicates absence of detectable T.

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gondii antibodies. However, a negative test result does not preclude the possibility of exposure
to or infection with T. gondii.
2.2 In addition to the presence of the C line, if only the G line is developed, the test indicates the
4. A negative or non-reactive result can occur if the quantity of the anti-T. gondii IgG or IgM
presence of IgG anti-T. gondii. The result is IgG anti-T. gondii positive or reactive and IgM anti-
present in the specimen is below the detection limits of the assay or the antibodies that are
T. gondii negative or non-reactive.
detected are not present during the stage of the disease in which a sample is collected.
5. The OnSite Toxo IgG/IgM Combo Rapid Test has not been validated on specimens from

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neonates.
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6. Infection may progress rapidly. If the symptom persists, while the result from OnSite Toxo
IgG/IgM Combo Rapid Test is negative or non-reactive, it is recommended to test with an
alternative test method.
2.3 In addition to the presence of the C line, if both the M and the G lines develop, the test indicates 7. Some specimens containing unusually high titers of heterophile antibodies or rheumatoid factor
the presence of IgM anti-T. gondii and IgG anti-T. gondii. The result is IgM anti-T. gondii and may affect expected results.
IgG anti-T. gondii positive or reactive. 8. The results obtained with this test should only be interpreted in conjunction with other diagnostic

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procedures and clinical findings.

REFERENCES
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1. Krick JA and Remington JS: Toxoplasmosis in the adult: An overview. New Eng J Med 1978.
298:550-3.
2. Anderson SE and Remington JS: The diagnosis of toxoplasmosis. So Med J 1975. 68:1433-43.
Samples with positive results should be confirmed with alternative testing method(s) and clinical
3. Rodrigues JP, Frei F, Navarro IT, et al. Seroepidemiological analysis of toxoplasmosis in
findings before a diagnosis is made.
en
college students. J Venom Anim Toxins incl Trop Dis 2015. 21:1.
3. INVALID: If no C line develops, the assay is invalid regardless of any color development on the 4. Montoya JG. Laboratory diagnosis of Toxoplasma gondii infection and toxoplasmosis. J Infect
test lines (M and G) as indicated below. Repeat the assay with a new device. Dis 2002.185(Suppl 1):S73–82.
5. Ramírez MLG, Alvarado VV, Gutierrez GV, et al. Prevalence of IgG and IgM anti-toxoplasma
antibodies in patients with HIV and acquired immunodeficiency syndrome (AIDS). Rev Soc Bras
S S
Med Trop 1997. 30(6):465-7.
6. Kravetz JD, Federman DG. Toxoplasmosis in pregnancy. Am J Med 2005. 118(3):212–216.
7. Calderaro A, Piccolo G, Peruzzi S, et al. Evaluation of Toxoplasma gondii Immunoglobulin G
(IgG) and IgM assays incorporating the new Vidia analyzer system. Clin Vaccine Immunol 2008.
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15(7):1076-9.
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Index of Symbols
Consult For in vitro
instructions for use diagnostic use only Use by
PERFORMANCE CHARACTERISTICS
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Tests per kit

1. Analytical Sensitivity of IgG Detection Catalog # Lot Number N
Twenty groups of matrix were spiked with the WHO International Standard Anti-Toxoplasma
Serum Ig (TOXM) concentrations of 0.625, 1.25, 2.5, 5, and 10 IU/mL. The specimens were run
Store between 2-30°C Do not reuse
on the OnSite Toxo IgG/IgM Combo Rapid Test. Defined as the 95% detection level, the limit of
detection or sensitivity for the OnSite Toxo IgG/IgM Combo Rapid Test IgG test line is 2.5 Manufacturer Date of manufacture
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IU/mL.

IgG (IU/mL) 0.625 1.25 2.5 5 10

Number Positive 0 11 19 20 20
SIMPLIFYING DIAGNOSTICS

CTK Biotech, Inc.

Number Negative 20 9 1 0 0
10110 Mesa Rim Road
2. Accuracy of IgG Detection San Diego, CA 92121, USA
Tel: 858-457-8698
A total of 237 clinical specimens were collected and tested on the OnSite Toxo IgG/IgM Combo
Fax: 858-535-1739
Rapid Test and by commercial ELISA. Comparison for all subjects showed 94.9% overall
E-mail: [email protected]
agreement for the IgG test line.
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PI-R0234C Rev. F
3. Accuracy of IgM Detection
Date released: 2016-07-29
A total of 231 clinical specimens were collected and tested on the OnSite Toxo IgG/IgM Combo English version
Rapid Test and by commercial ELISA. Comparison for all subjects showed 97.8% overall
agreement for the IgM test line. For Export Only. Not For Resale In The USA.

4. Performance on BBI Anti-T. gondii Mixed Titer Performance Panel

The performance of the OnSite Toxo IgG/IgM Combo Rapid Test was evaluated using the BBI
(Boston Biomedical Inc) Anti-T. gondii Mixed Titer Performance Panel (PTT202) and compared
with three commercial immunoassays. The comparison is shown in the following table:

BBI Reference Abbott bioMerieux Diasorin OnSite Toxo IgG/IgM

Panel (PTT202) ARCHITECT VIDAS LIAISON Combo Rapid Test
IgG Positive 20 20 20 20
IgG Negative 1 1 1 1
IgM Positive 8 7 6 6
IgM Negative 13 14 15 15

5. Cross-Reactivity
No false positive anti-T. gondii IgG and IgM results were observed on 3-14 specimens from the
following disease states or special conditions, respectively:

CMV Dengue HAV HBV HCV

HIV HSV-1 HSV-2 hCG H. pylori
Malaria TB T. pallidum Rubella ANA
HAMA RF (up to 8400 IU/mL)

Copyright 2016 by CTK Biotech, Inc.