Blood banking Discussion 1

DONOR SELECTION
Donor- less harm.
Purpose
Recipient- will have benefits.

Process:

a. Donor ID
b. Donor medical history
c. Medical assessment (Physical exam)
d. Hematocrit, hemoglobin and blood typing
e. Component preparation

a. Donor ID- demographical data (also includes contact number/ cp number, email)
- Confirmation (Id card) or vouched.

** NVBSP- NATIONAL VOLUNTARY BLOOD SERVICE PROGRAM.

b. Medical history- includes donor assessment.

- Self-filled up forms (DHQ).
1. Are you feeling well today?
2. Are you taking medications?
o Aspirin- can donate whole blood
- If the patient will use it for platelets, then the donor will be deferred for 72
hours or 3 days.
o NSAIDS (paracetamol)
- Can donate whole blood, for platelets, donor will be deferred for 24 hours or 1
day.
- If donor uses it for fever, then he/she cannot donate blood for that time or day.
o Anti-hypertensive medications
- Can donate whole blood if BP is controlled.
o Diabetic medications
 Insulin- defer permanently
- Because insulin is a subcutaneous injection which causes transient
bacteremia.
 Oral hypoglycemic agents
- Refer to physician.
- If controlled whole blood can be collected, if not, defer donor.
o Herbals/ supplements/ vitamins
- Ask why they take those drugs.
- If there are allergies- defer as long as the donor is ingesting or taking the
supplements + 72 hours (3days).
 - If there’s no allergies- can take blood.
o Glutathione
 Injection- defer
 Oral- as long as the indication of the GSM is not cancer, we can take
blood.
o Retinoic acid (high doses of Vitamin E)
- topical, problem is it may still enter the system because of the small vessels
on the skin which can cause birth defects specially if taken orally defer for 4 weeks or 1
month.
- If used as maintenance because of acute promyelocytic leukemia-
permanent deferral.
o Allergy medications
- As long as taken orally plus no signs and symptoms- can be a donor
(deferral will not be very long).
- Injection – moderate- severe reactions, defer for 1 whole year.
- Allergic shock/ anaphylactic shock- permanent deferral.
o Iron supplements
- Ask why the donor is taking the drug.
- Anemia- iron deficiency- deferred until stop medication.
- Post operation/ delivery/ other reasons aside from iron deficient will be
based on the discretion of the physician (deferred for at least 1 year).
- Thalassemia/ chronic kidney disease- permanent deferral.

** stable – 125 min. for hemoglobin patients.

o Antibiotics
- Antibiotic is not the reason for deferral.
- What is the infection?
o Anti- cancer medications?
- Permanent deferral.
o Vaccines
 Live attenuated- Mumps, measles, oral polio vaccines, anti-rabies (prophylaxis-
post exposure), BCG, anti-small pox, yellow fever, typhoid fever (oral)= deferred
for 1 month except for rabies which will be deferred for 1 year.
 Killed/ toxoids- influenza vaccines, tetanus toxoids, pneumococcal vacs, HIB,
anthrax, cholera, diphtheria, hepa B, hepa A=can donate blood anytime except
for hepa B which will be deferred for at least 1 week.

**HIB given specially to HCW working in microbio laboratory.

***Rubella deferred for 8 weeks (2 months).

***MMR- combination vaccines deferred for 2 months.

***MMR, BT deferred for 1 month

 Anti- covid 19 -DOH
o SINOVAC- inactivated- deferred for 28 days.
o JANSSEN (J&J), ASTRAZENICA, SPUTNIC – viral vector based- defer for 28 days
o PFEIZER, MODERNA- mRNA bases- accept donor anytime.
o BHARRAT BIOTECH- deferred for 28 days
o NOVAVAX- subunit- anytime.
o CODAGENIX- inhaled not injected; live attenuated virus-deferred for 28 days.
o No signs and symptoms- deferred for 28 days.
Dengue vaccines- deferred 1 month.
HPV (human papilloma virus) - to prevent cervical cancer (girls)
- Can also prevent renal cancer in males.
Blood transfusion, dental extraction, tattoo, body piercing, acupuncture, sexual contact of high-
risk individual, living with people with hepa B, imprisoned, relative with Creutzfeldt Jacob
disease/ mad coco (prion disease) = all will be deferred for 1 year.

** to avoid HIV, HB, HC and syphilis there is a window period.

Last menstrual period

 Without pregnancy- don’t get blood from them
 With menstruation- defer until the last day of menstruation.
Indefinite deferral – menorrhagia- too much blood
- Metrorrhagia- too often to have mens
- Menometrorrhagia- combination of the two.
 Pregnancy- defer for 12 months after delivery or 3 months after weaning whichever is longer.
 Abortion as long as bleeding stops, can be a donor.
*Philippines {raspa/ D&c}- defer for 1 year.
**No d&c, patient is given rest, deferred for 6 weeks.
 Blood donation- can donate 3 months after past donations.
 4X for 1 year blood donation- provide whole blood donation.
 Greater or equal to 300ml of blood= deferred for 12 weeks/ 3 months.
 Less than 300 ml, deferred for 6 weeks.
 Special case: males may be allowed to donate after 6-8 weeks if emergency. Also, if they pass
the DHQ, PE and HGB.
 Residence- UK – OJD- permanently deferred.
- Europe – 5 years- indefinitely.
- Sub Saharan Africa – 5 years- indefinitely. (Nigeria, Niger, Congo, Chad,
Cameroon)
- Ph malaria endemic areas- Kalinga and Abra (probationary period), Palawan.
 Recipient of clotting factors- coming from human blood.
- Deferred for at least 1 year but if multiple times recipient will have permanent deferral.
 Patient positive for HIV, HB and syphilis- permanent deferral and will undergo counselling.
 Hepatitis – needs to know the history.
 Hep. A (Contagious) – oral route
-Acute
 -Less than 10 years old when he/she acquired hep. A- can be
accepted as donor.
 Not Hep. A deferred indefinitely- HBsAg, anti HBS, anti HCV.
o Acute hepatic disease, chronic liver disease- HCCA.
 Malaria (+) – deferred permanently.
 Malaria like illness-ask if it is more then 3 years, if more than 3 years= accept as donor, if
less than 3 years defer still until it is already 3 years.
 Warts, syphilis, herpes-deferred permanently.
 Cancer (basal cell CA, Sg cell CA, Cervical cancer in-site) – deferred permanently.
 History heart attack (AMI), CHF, Chronic bronchitis (acute exacerbation), bronchial asthma –
permanently deferred.

c. Physical exam
o Age
 16-60 years old
 16-17 needs legal guardian
 >60 can still donate provided that they are regular donors/ donors for 1 year.
 70 years old refer to MD.
o Weight
 Minimum 50 kg (10.5 ml/kg).
 405-495 ml (450+45 ml >63 ml preservative)
 350 ml blood- low volume blood unit.
- Not used to prepare for platelet concentrate.
- Ffp
- Prbc- for pediatric patients.

If donor is 40 kg, calculate.

40 kg X 10.5/1kg = 420 ml

450 ml/420 ml= 63ml preservative

X= 58.8 ml

63-58= 5 ml.

**if donor is 38kg, no need to remove or lessen the preservative.

PHYSICAL EXAM PROPER

Blood pressure
 Systolic ≥160
 Diastolic ≥100

*if systolic is ≥180 refer to physician.

 Pulse rate
 60-100 beats per minute- 60 seconds
 50- athletic or physically active person (can still be a donor).
 <50 deferred.
o Bloodletting is 15 minutes
o Check for:
 Temperature- should not be febrile.
 Consciousness
 Skin
 Eyes/ sclera/ finger nails: if anemic defer.
 Listen to heart and lungs
 Look at the feet (lesions): try to look between the big toe and the 2 nd toe and the side of the
foot. Some donor injects drugs from these areas.
d. Hematocrit, hemoglobin and blood typing
 Hematocrit- microcapillary centrifugation method.
- Males have relatively higher value than female.
- Male: 0.40
- Female: 0.38
- Note: hgb is the most important.
 Hemoglobin – copper sulfate method (finger prick) 125g/dl.
- Immunochromatography
- Male: 135g/dl
- Female: 125g/dl
 Blood typing- slide method for mobile blood banking.
- Blood banking: tube method
e. Component preparation
 Whole blood
1. Processing
- Repeat blood typing (tube method).
- Set aside tubings
- Screening for TTIs
- HIV 1 and 2, HCV, HBV, malaria and syphilis.
- Records management
2. Component separation
- Single blood bag- limited (whole blood, packed red blood cells)
- Double set- whole blood, prbc, ffp
- Triple set system- whole blood, pRBC, ppfpp, platelet concentrate (room temp not >25C).
- *both single and double set system are stored on ref temp which is 4-5 degrees Celsius.

**whole blood use cryocentrifuge.

SCREENING FOR TRANSMITTED DISEASES

Required Test
 Syphilis
 HBsAg
 HIV-1,2antigen
 Antibodies to HIV-1,HIV-2,HBC and HCV
 Anti-HTLV-I-II
 Malaria

Screening for Infectious Diseases at PGH Blood Bank

 HEPATITIS B (HBsAg)
 HEPATITIS C (anti-HCV)
 HIV (HIV-1 antigen,anti-HIV-1 and anti-HIV-2)
 SYPHILIS (RPR)
 MALARIA (thick and thin smears)

HEPATITIS
 Hepatitis lead to transfusion is almost exclusively caused by viruses

1. Hepatitis viruses A to B
2. CMV
3. EBV
4. newly described putative hepatitis viruses such as GBV-C,TTV and SEN-V

HEPATITIS B
 Current screening immunoassays detect approximately 0.2 to 0.7 ng/ml HBsAg
 Electrochemiluminescence immunoassay –in vitro qualitative determination of HBsAg in human
serum and plasma

 Test principle sandwhich principle

 HBsAg specific antibody + sample with Ag + HBsAg specific antibody labeled with
ruthenium complex
 Add streptavidin-coated microparticles -> complex becomes bound to the solid phase
 Microparticles are magnetically captured onto the surface of the electrode
 Application of voltage-> induces Chemiluminescent emission measured by a
photomultiplier

False positive - send samples for confirmatory testing

  False negative results: happens because of antigen concentration below the detection limit of the
assay and the lack of reactivity of the antigens to the antibodies used in the assay
 100% sensitivity
 99.7% initially reactive specificity
 100% repeatedly reactive specificity

HEPATITIS C
 Test for antibodies to HCV – enzyme immunoassays (EIAs) using recombinant antigens of HCV
coated on a solid phase as the capture reagent
 Principle: Two step indirect solid phase enzyme immunoassay

 False negative results: antibody concentration may be beneath the detection limit of the assay
(such as in early infection ), immunosuppressed patients, patients antibodies do not react with the
antigens used in the assay
 100% sensitivity and 99.7% specificity
 Repeatedly reactive EIA’s -.> confirmed by recombinant immunoblot assays (RIBA)

HIV
 Etiologic agents of AIDS
1. HIV-1 (US,Europe and central Africa)
2. HIV-2(West Africa)
 Retrovirus that preferentially infects CD4-positive T lymphocytes (helper T cells ) in lymph
nodes and other lymph node tissue
 P24 antigen screening reduces the window period by 6 days
  HIV combi EIA: qualitative detection of HIV-1 p 24 antigen and antibodies to anti-HIV 1 group
M and group O and HIV-2 in human serum and plasma
 Test principle: double antigen immunoassay for the detection of total antibodies to HIV-1 and
HIV-2, combined with a sandwich assay for the detection of HIV-1 p24 antigen.

Explanation:
Reagent antibodies will be incubated along with again with the microbid, this microbid will now
have combination with these antibodies and it will also have antigens. Incubate them together and then
after incubator add the second set of reagent which is the solid waste because they are now attached to the
bottom and sides of wheel. Wash and then you will add buffer to make the connection permanent. The
buffer there will change the shape of the proteins so that their connections will be more permanent , wash
it and you detect it.
 False positive result: contamination during measurement of HIV-1 p24 and anti-HIV 1 and anti-
HIV 1
 Specificity = 99.85%
 Sensitivity = 100%
 2004=30 unconfirmed cases

SYPHILIS
 Caused by the spirochete Treponema pallidum
 Syphilis transmission by transfusion is not effectively prevented by subjecting the donor blood to
standard serologic tests for syphilis (STS) because seroconversion often occurs after the phase of
spirochetemia
 RPR card test is a qualitative and semi-quantitative non-treponemal flocculation test for the
determination of reagin antibodies in human serum

False positive results:

1. samples from individuals with history of drug abuse
2. individuals with diseases such as lupus, erythematous, mononucleosis,leprosy,viral,pneumona
3. after small pox vaccination

 Interferences: contaminated lipemic or grossly hemolysed specimens should not be used because
of the possibility of non-specific reactions
 Othe rmethods used: detection of IgG and IgM for treponema pallidum,ELISA/EIA
 2004-39 unconfirmed cases

MALARIA
 Caused by several species of the intraerythrocytic protozoan genus plasmodium
 No practical serologic test to detect transmissible malaria in asymptomatic donors travelling to
endemic areas are often deferred from donating blood for 1 year
 Screening: thick and thin smears

HUMAN T0CELL LYMPHOTROPIC VIRUSES

A. HTLV type 1 – associated with a dults T-cel lymphoma/leukemia
 Assoc with neurologic condition HTLV-associated myelopathy (HAM)
 Donor screening began in the uS in late 1988

B. HTLV type II – at least 6-\0% homology of genetic sequence to those of HTLV-I

 Associated with HAM

 Transmission:contact with infected viable lymphocytes

 EIA: donor screening for anti-HTLV I/II
 Risk of transmission in the United States – 1 in 641,000
 No licenses confirmatory test for HTLV

INFECTIOUS DISEASES NOT ROUTINELY SCREENED

A. CYTOMEGALOVIRUS
 Member of human herpes virus family
  Immunocompetent persons may be asymptomatic
 Causes serious morbidity and mortality in premature infants, recepients of organ, marrow or
peripheral blood progenitor cell transplants; and AIDS patients
 Leukocyte removal to 5x to 106 per component significantly reduce, if not prevent, post
transfusion CMV

B. EBV
 Causes most cases of infectious mononucleosis;closely associated with endemic form of Burkitt’s
lymphoma and nasopharyngeal carcinoma
 Rare cause of post transfusion hepatiris
 Prevention:use of leukocycte reduced blood products

C. Parvovirus V19
 Cause of erythema infectiosum or 5th disease
 Infect and lyse red cell progenitors in the marrow -> sudden and severe anemia of patients with
chronic hemolytic disorders
 Infection during pregnancy-> spontaneous abortion, fetal malformation and hydrops from severe
anemia and circulatory failure
 Screening- not a high priority because of the benign and/or transient nature of most parvovirus
infection and extreme rarity of reports of parvovirus 19 transmission by individual components
1. Donor History
2. Physical examination
3. Serologic screening

REDUCING THE RISK OF INFECTIOUS DISEASE TRANSMISSION

1. Inactivation or destruction of agents in derivatives or plasma products
a. Application of organic solvents or detergents -> inactjvates viruses with a lipid-
containing envelope (HIV,HBV,HCV,HTLV)
b. Heat treatment
c. Photochemicals (psoralen activated by UV A light used in inactivation of platelets and
plasma)
2. Use of leukocyte-reduced blood products
3. Proper screening of all blood products

ABO DISCREPANCIES & OTHER PROBLEMS

DISCREPANCIES
 A discrepancy occurs when the red cell testing does NOT match the serum testing results
 In other words, the forward does NOT match the reverse (Not advisable to use slide method dahil
forward lang yon. Nirerequire ng DOH is tube method para makita natin kung totoong yon ang
blood type of the donor) It help us in the proper selection of the blood that we will release for
blood transfusion.
WHY?
 Reaction strengths could be weaker than expected
 Some reactions may be missing in the reverse or forward typings
 Extra reactions may occur

WHAT DO YOU DO?

 Identify the problem
 Most of the time, the problem is technical
•Mislabeled tube
•Failure to add reagent
•Either repeat test on same sample, request a new sample, or wash cells
 Other times, there is a real discrepancy due to problems with the patient’s red cells or serum

DISCREPANCY?
 If a real discrepancy is encountered, results must be recorded
 However, the interpretation is delayed until the discrepancies RESOLVED.
(Kumbaga huwag kang magpapalabas ng blood type until the discrepancy is resolved. Once
nagpalabas ka at kailangan pala ng pasyente ng dugo magoorder na si Doc ng cross matching
form mas marami ka nang paperworks na hahabulin)
TECHNICAL ERRORS
Clerical errors
 Mislabeled tubes
 Patient misidentification
 Inaccurate interpretations recorded
  Transcription error
 Computer and entry error

Reagent or equipment problems

 Using expired reagents
 Using an uncalibrated centrifuge
 contaminated or hemolyzed reagents
 Incorrect storage temperatures

Procedural errors
 Reagents that added
 Manufacturers directions not followed
 RBC suspensions incorrect concentration
 Cell buttons not resuspended before grading agglutination

CLOTTING DEFICIENCIES

 Serum that does not class maybe due to:

 Low platelet counts
 Anticoagulant therapy (Heparin, Aspirin, etc)
 Factor deficiencies
 Serum that does not clot completely before testing is prone to developing fibrin clots that may
mimic agglutination
 Thrombin can be added to serum to activate clot formation
 OR, tubes containing EDTA can be used (CPDA1) -Donor units

Serum may not be a good sample for blood typing kaya most of the time ang ginagamit natin is EDTA.
CONTAMINATED SAMPLES OR REAGENTS

 Sample contamination (when blood is more than 48 hours)

 Microbial growth ( they will eat antigens of RBCs. Remember that yhe antigens in RBCs
contains sugar so they like to eat sugar, now the bacteria can degrade your glycoproteins.
They like to eat B antigens for some reasons)
 Reagent contamination (kakadotdot and kakabukas ng blood typing sera nagkaroon na ng
bacteria)
 Bacterial growth causes cloudy or discolored appearance… do not use if you see this! Main
problem: Fungi
 Reagents contaminated with other reagents (don't touch side of tube when dispensing)
 Saline should be changed regularly Fungal hyphae

EQUIPMENT PROBLEMS

 Routine maintenance should be performed on a regular basis (daily,weekly, etc)

  Keep instruments like centrifuges, thermometers and timers calibrated
 Uncalibrated serofuges can cause false results

Quality management. Make sure all the centrifuges are maintained tamang speed, level, timers calibrated
also, thermometers, lalong lalo na pag mag ko crossmatch na kayo.
HEMOLYSIS (Hemolysis is not an issue usually for your blood units. Hemolysis is more a problem
for your patients when you encounter a difficult extraction)

 Detected in serum or plasma after centrifugation (red)

 Important if not documented
 Can result from:
 If it is not due to difficult extraction, it might be due to the presence of Complement
binding to your RBCs
Who are these complements that like to cause hemolysis in our blood sample?
Anti-A, anti-B, anti-H, and anti-Lea because a lot of them can work at room
temperature
 Bacterial contamination do not use blood in prolonged storage

ABO DISCREPANCIES

 Problems with RBCs

 Weak- reacting/ Missing antigens
 Extra antigens
 Mixed field reactions (when looking at the microscope and assessing coagulation may areas
na ang ganda ng clumping of red cells tapos kalat kalat dito, tapos may area na naman na
maganda ang clumping niya)
 Problems with SERUM
 Weak- reacting/ Missing antibodies
 Extra antibodies
This is an illustration of different causes of Discrepancies of Forward typing and in Reverse typing.
For your Forward typing the problem is usually in your RBC antigens. If there is a problem in RBC
antigens and there is missing then it might be because of these subgroups of your type A or type B, or it
might also be because of cancer.
Extra reaction to RBC antigen can be in acquired B phenotype, B(A) phenotype or it can be due to
abnormal proteins in your plasma abnormal proteins can now cause rouleaux formation
Mixed field can be due to transfusions using blood group O. What does it mean? It’s the 2 nd or 3rd time na
ni patient na mabigyan ng dugo so ipapacrossmatch ulit . Noong una pala siyang na transfused hindi pala
O type kasi emergency. Example: Meron kang patient which is type B dahil emergency instead na B ang
itataransfused type O tapos ung pagdudugo niya is tuloy tuloy pa rin. After 2 daya kailangan niya ulit ng
dugo so since hindi na emergency ipupunta na sa bloodbank yung dugk niya properly for proper typing
and proper crossmatching. Nong nagba bloodtype ka na dun mo nakita na may mixed field. Probably
because our anti-sera is reacting with the B cells of your ….. and at the same time you still have O cells
from the previous donation. May mga areas na nagka clump yung mga B cells ng patient mismo and may
mga areas na yung O nakakatransfused lang ay nandon na tumatambay lang walanv nagreact sa kanila.
Pwedeng type A din yan for as long as bina blood type mo siya yun pala natransfused san na siya ng type
O dati na wala pang 72 hours or 24 hours. The same thing applies for patient who are given Bone marrow
transplant kase ang main na babol natin sa mga patient na may bone marrow transplant is the HLA
compatibility and also ABO compatibility. Sana mag match yong donor and recipient ng HLA antigens
and magmatch din sana sila sa ABO antigens. But sometimes when patient have bone marrow transplant
there is a period of time na kung saan yung bone marrow transplant nag aadjust palang hindi niya
mailabas yjng buong complement ng ABO antigens so magkakaroon ka rin ng mixed field agglutination.
Mixed field go back and ask the nurse in the ward Ma’am apay tatransfused san yu nji patient ana
indication nanatransfuse san datin ana intransfused da idi type O ba? Or Ma’am adda ba leukemia ni
patient sinuktan da ba tay bone marrow na? Mag ask kapag mah mixed field agglutination. Most of the
time history mareresolve mo na yan.
For Reverse typing there is something usually wrong in the Serum it has something to do with
antibodies. They might be weak and you see them in this cases Young Elderly Immunocompromised.
Ask the nurse Ma’am detuy patient ada ba AIDS na? Apay inadmit yo Ma’am sobra ba SLE na inkabil yo
request form? Adda ba tumtumaren? Adda ba cancer ni patient? Immunocompromised you may be able to
see weak antibodies. Extra it can be Cold Autoantibody , Cold Alloantibody, there might be again normal
proteins jn your blood and it will now caused Rouleaux formation and you might have Anti-A1 kasi yung
patient pala is subgroup A2 so magkakaroon siya ng Anti -A1. Kung yung donor subgroup A2
magkakaroon siya ng Anti-A1 yun na yun ang magiging rason kung bakit meron kang extra reaction sa
reverse typing.
Take Note: Rouleaux can be seen in both Forward and Reverse typing.
Remember:
1. Rouleaux should be very sure na rouleaux yung nakikita ninyo. Dapat stack of coins
arrangements. (linear)
2. Rouleaux formation is due to abnormal protein in the blood and a lot of cases in this has related
to patient undergoing hemodialysis.
If you see rouleaux formation, check the record check kung tama ang tube labels check kung
tama mga machines ninyo centrifuge lalo na and then the next step for probably kapag ginagawa
mo yun iyong staff niyo naman magtatankng na sa nurse station. Ma’am mag hemodialysis ba si
patient.

COMPATIBILITY TESTING PROCEDURES

 (Consensus on Pre-Transfusion Practices, 2001)

Usually done in the Philippines:

 ABO Testing
 Rh Testing
 Historical Record Check

While do not routinely do the ff:

o Screening for unexpected alloantibodies – see during ABO testing
o Alloantibody identification
o Management of previously identified alloantibodies – see during Crossmatch,
it is not screening anymore because you detect it already.

NOTE:
 If the donor is a known carrier of alloantibodies, then it should not be able to
suggest on how to manage them.
 The main part of Compatibility testing is the CROSSMATCH.

Purpose of Compatibility Testing:

 For the safe blood transfusion therapy.

LIMITATIONS:
o Cannot guarantee normal survival of transfused red blood cells in the
recipient’s circulation.
o Cannot guarantee that no adverse response to transfusion will occur.

PRE-REQUISITES TO PROPER COMPATIBILY TESTING

1. Positive Recipient Identification

 Misidentification of the recipient: most common cause of transfusion error.
 Patient’s Wristband ID should always be compared with the blood request
form.
o Nameplates on the walls or bad labels should never be used to verify
identity.
 If coherent: ask patient to state full name and to spell it out.
o Never offer a name (e.g., “Are you Mr. Dela Cruz?”)
 If incoherent or very young: reliable individual who knows the patient must
confirm the identity.
 If patient’s identity is not known: temporary designation until proper
identification.

2. Proper Collection of Patient Samples

 Hemolysis should be avoided
 10mL is usually sufficient
  Most prefer Serum (Red Top), although Plasma (Violet Top) can also be
used.
o Serum: has no fibrin and does not inactivate complements.
 Tube must be labeled properly and legibly before leaving the bedside.
 Do not collect at IV-line sites
o If IV line is at ante-cubital fossa: may collect blood from veins BELOW
the IV lines.
o If unavoidable: stop the infusion for 5-10 mins, discard first 10mL of
extracted blood, then collect sample for testing.
 All discrepancies must be resolved before accepting the sample at the
laboratory
o In case of doubt: draw new sample
 Testing should be done as soon as possible after collection.

3. Proper Collection of Donor Samples

 Samples for donor testing should be collected at the same time as the unit of
donated blood, and should bear the same identification code.
 Samples for compatibility testing can be obtained from the tubing (do not
puncture the blood bag) of the blood unit.

COMPATIBILITY TESTING PROCEDURES: ABO GROUPING

 The most critical pre-transfusion Serologic Test is ABO GROUPING
 Either slide or tube method
 Do forward and reverse grouping (always)
o Discrepancies between forward and reverse grouping: Do additional
Testing

COMPATIBILITY TESTING PROCEDURES: Rh GROUPING

 Tube or slide method
o Do not use slide because we are dealing with Rh grouping.
o Rh Grouping used as Rapid Identification of the Rh type.
 Weak D testing: may not necessary for recipients (rarity of partial D group)
o If blood group cannot be satisfactorily determined and immediate
transfusion is required: Give Group O Rh (-) pRBC.
NOTE: Never give type O whole blood for emergencies, but use type O packed red
cells for emergencies. Because whole blood has PLASMA, Plasma of Group O has
Anti-A, Anti-B, Anti-AB it is not good for the recipient.

GROUP O is the universal donor of red cells, not universal donor of whole blood.

COMPATIBILITY TESTING PROCEDURES: ANTIBODY SCREENING

Objective: to detect as many clinically significant unexpected antibodies as possible.

 Antibodies that are:

o Reactive at 37˚C AND/OR
o Reactive in the Anti-Human Globulin Test, and
 o Known to have caused a transfusion reaction or unacceptability short survival
of transfused red cells.
 Usual antibodies: it is most often associated with Hemolytic Transfusion
Reactions
o ABO
o Rh
o Kell
o Duffy
o S,s,U
o P
 Especially for multiple transfusion recipients.
o The more frequently a patient is exposed to foreign RBC antigens, the more
likely that a patient will produce unexpected alloantibodies.
 Important for:
o Selection of donor RBC’s that will have the best survival rate in the patient’s
circulation
o Reducing the risk of hemolytic transfusion reaction

 All antibodies encountered must be identified to:

o Determine clinical significance
o Allow logical decision to be made whether there is a need to select antigen-
negative units for transfusion.
 A semi-automated antibody screener (Capture-R Ready Screen I and II)
 Principle of the solid-phase antibody capture procedure used by Capture-R Ready
Screen I and II.
 Grading of results in the Capture-R Ready Screen I and II. It is start with 0, 2+, 4+.
You need to observe for the agglutination and hemolysis.
 Master list of antibodies detectable using Capture-R Ready Screen I and II.
o IgM antibodies: M, N, Le+, P1
 Usually react on immediate spin
o IgG antibodies: Kell, Rh, Kidd, Duffy
 Usually react in the AHG phase

 TYPE and SCREEN

o ABO grouping + Rh grouping + Antibody Screening
o Unexpensive procedure

COMPATIBILITY TESTING PROCEDURES: CROSSMATCH TESTING

Value:
 It is final check of ABO compatibility between donor and patient.
 It may detect the presence of an antibody in the patient’s serum that will react with
antigens on the recipient’s RBCs but was not detected in the antibody screening
because the corresponding antigen was lacking from the screening cells.

Serologic Crossmatch Testing

Traditionally classified as either:
 Major X-Match
o Recipient serum + donor cells (RS-DC)
o Detects antibodies against the donor’s red cells
 Minor X-Match
o Recipient Cells + Donor Serum (RC-DS)
o Detects antibodies against the recipient’s red cells.
 Elimination of the Minor X-Match
o Presence of low-incidence antibodies in donor’s plasma would not cause
transfusion reaction since it would be diluted in the recipient’s plasma.

TRADITIONAL PHASES
o Saline phase/Immediate Spin (IS)
o 37 ˚C incubation with low ionic solution (LISS)
 a.k.a LISS Phase “Bovine Albumin” Phase
o Anti-human globulin (AHG) Phase

 Immediate Spin Phase

o Designed to detect ABO incompatibilities between the donor RBC and px
serum.
o It detects usually cold antibodies

 LISS PHASE
 Low Ionic Strength Solution (LISS)
o Includes 22% or 30% bovine albumin, polybrene and polyethylene glycol.
o Enhances sensitivity by increasing the rate of antibody uptake and shortens
incubation time.

 Anti-human globulin (AHG) Phase

o An indirect antiglobulin (Coomb’s) test
o Demonstrates the presence pf antibody or complement which results in
the in-vitro coating of RBC.

CROSSMATCH TESTING:
USE OF AUTOCONTROL
 Patient’s own serum + own RBC and tested similarly as the major crossmatch
procedure
 Results can help clarify possible explanations for positive results in crossmatch.
 A NEGATIVE result effectively rules out the presence of autoantibodies.
 No longer required in the current AABB Standard.

NOTE: it is always a good practice to include an auto control every time you perform
crossmatch.
COMPATIBILITY TESTING PROCEDURES: ANTIBODY SCREEN AND
CROSSMATCH
 Originally: Crossmatch then Antibody Screening.
 Current standards: Antibody screening then crossmatch
o Almost 99% of clinically significant unexpected antibodies can be already be
detected in antibody screening.
o Crossmatch can be shortened (Abbreviated crossmatch)
 “Type and Screen” then IS phase only
 Studies: a safe and effective way of pretransfusion testing.
 Abbreviation Crossmatch
 Problems
o The Immediate Spin Phase does not detect all ABO incompatibilities
o False reactions may be seen:
 In the presence of other immediate spin reacting antibodies (e.g., Anti-I)
 In patients with hyperimmune ABO antibodies
 When procedure is not performed correctly
 When rouleaux occurs
 When infant’s specimens are tested
 Addition of EDTA to the IS solution: can eliminate some false positive
reactions.
 Should not be done if antibody testing showed presence of a clinically
significant alloantibody.

GUIDELINES IN THE INTERPRETATION OF RESULTS

 All tubes or gel cards should be labeled carefully at all stages of the procedure.
 Results should be read against a white or lighted background.
 If needed, a magnifying lens can ben used to facilitate reading.
 Button of RBCs after centrifugation should be gently re-suspend
o Violent shaking may cause false negative readings.
 Grading of agglutination reactions should be uniform among the staff
 All results should be recorded immediately
 If incompatibility is found, additional testing should be performed and records should
clearly show the location of the results of the follow-up studies
.
CAUSE OF POSITIVE RESULTS IN THE SEROLOGIC CROSSMATCH
 An antibody in the patient’s serum reacting with the corresponding antigens
on donor RBC:
o The alloantibody should be detected and identified in the antibody screen.
o After identification, crossmatch is repeated using units negative for the
corresponding antigen.
o If unable to identify, consult a reference laboratory.
 Solution: Auto-absorption
 Prior coating of the donor RBC with protein, resulting in a positive AHG
Phase
o Do a Direct Antiglobulin (Coomb’s) Test on the donor’s red cell
o Positive DAT  donor’s cell is already coated with immunoglobulins and/or
complement.
o Donor units positive DAT should not be used. (Do not give to others)
  Abnormalities in patient’s serum:
o Imbalance in the normal A/G ratio
 Occurs in multiple myeloma, macroglobulinemia
 Leads to rouleaux formations, which mimics true agglutination  verified
microscopically
 Rouleaux is seen in LISS phase but do not persist in AHG phase
 Solution: Saline Replacement Technique
o Presence of high molecular weight dextran’s or other plasma expanders.
 Solution: Saline Replacement Technique
o Antibody against additives in the albumin reagents.
 Solution: Use Albumin reagents without caprylate

COMPUTER CROSSMATCH
 A.K.A. Electronic Crossmatch
 Compares recent ABO serologic results and interpretations on file for both the donor
and the recipient being matched
 Determines compatibility based on the comparison.
ADVANTAGES:
 Increased annual savings
 Reduced sample requirements
 Reduced handling of biologic material
 Elimination of false reactions encountered in the IS phase
 Safer than the IS phase
 AABB Standards for the use of computer crossmatch
o Can only be used for the purpose of detecting an ABO incompatibility
o Current antibody screen should be negative and there must be no history of such
antibodies
o At least two concordant patient ABO-Rh types must be on file
 Computer System:
o Must have logic to notify the user of the following:
 Discrepancies between donor ABO-Rh type on the unit label and those
determined by blood group confirmatory tests.
 ABO incompatibility between the recipient and donor unit.
o Must be validated

COMPATIBILITYU TESTING IN SPECIAL CIRCUMSTANCES

 Emergencies
o Strategies:
 Perform a shortened crossmatch procedure then release (usually only typing
then LISS phase) the blood unit
 Release a blood unit while the crossmatch procedure is being completed
(usually released after the IS phase).
  Release Type O Rh Negative packed red blood cell unit if there is extreme
emergency and there is no time to do blood typing.
o Tag labeled must be placed on the unit indicating that compatibility testing was
not completed before the release of the unit
o Physician must sign a release authorizing and accepting responsibility for using
blood products prior to completion of compatibility testing
o Any subsequent incompatibility result should be immediately reported to the
physician in charge.

 Transfusion of Non-Group Specific Blood

o E.g., Type O units were transfused to a Type A patient:
 Test patient’s serum for anti-A and anti-B prior to giving additional RBC
transfusion.
 If compatible with original blood type: give co-type blood (i.e., Type A Units)

 Compatibility Testing for Transfusion of Plasma Products

o Not required
o If large volume is required, may do IS phase crossmatch between recipient red
cells and donor plasma

 Intrauterine Transfusions
o If fetal blood type is known and there is no fetomaternal ABO and Rh
incompatibility: use fetal blood co-type unit.
o If fetal blood type is not known or if there is fetomaternal ABO/Rh
incompatibility: use type O Rh Negative unit
 For crossmatching: use Mother’s Serum
o Unexpected antibodies in the mother’s circulation might cross the
placenta (e.g., anti-K1 and anti JK)
o Blood for transfusion: as fresh as possible, no more than 7 days.

 Neonatal Transfusions (less than 4 months old)

o Should be compatible with maternal antibodies in the neonatal circulation
o Do ABO and Rh typing of mother and infant
 Forward typing only for infants
o Antibody testing: use maternal blood
 Infant’s blood is used only when there is no maternal specimen available or
when mother has clinically insignificant antibodies.
o The presence of clinically significant antibodies
 Cells lacking the corresponding antigen must be selected until the antibody is
no longer demonstrable in the infant’s serum
o Blood for transfusion: as fresh as possible, no more than 7 days.

 Massive Transfusions
o “Massive”: amount of blood transfused approaches or exceed the patient’s total
blood volume
 o Antibody in the patient’s serum may not be demonstrable because of dilution with
large volume of plasma and other fluids.
o If the patient is already known to have a clinically significant antibody, all units for
infusion should be tested for the corresponding antigen, if time permits.
o Physician may decide to shorten or eliminate compatibility testing after signing a
waiver or a release form for all units.

 Specimens with Prolonged Clotting Time

o Coagulation abnormality may be due to disease or medication (heparin).
o Poor clotting of blood  left-over fibrin during separation of serum  fibrin clot
formation during IS phase  false positive.
o Solution:
 Add thrombin (50U/mL) to serum/plasma (1mL) to induce clotting prior to
testing.
 Add protamine sulfate to blood to counteract effect of heparin.

 Autologous Transfusion
o Do ABO and Rh grouping
o No need for donor blood screening and antibody screen if blood is to be used
within the same facility.
o Antibody screening and crossmatch testing is optional if blood was not collected
from the same institution.
o Unit must be labeled “for autologous use only”