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Transcript of Blood Bank Practical
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The Osler I nstitu te
P}Chaffin (2/11/2013) Blood Bank Practical page 1
Blood Bank Practical D. Joe Chaffin, MD
Cedars-Sinai Medical Center, Los Angeles, CA
Outline
1. Specific Situations2. Calculations3. Antibody ID
I. Specific Situations in Transfusion MedicineA. Emergency transfusion/acute hemorrhage
1. Situations of acute blood loss where there may not betime for full pretransfusion workupsa. Need is more urgent for volume than for oxygen
carrying capacity; fluids more critical in early stages1) Historically, crystalloid was used for volume2) Excessive crystalloid can be harmful; has led to re-
emphasis on colloids instead b. Most common in trauma settings, and commonly leads
to massive transfusion (see below)c. Loss of over 30% of blood volume leads to significant
clinical consequences and increased mortality2. Priority : Dont harm anyone by doing something stupid!
a. High stress situations b. Staff must train and drill for these events ; its not the
time to be learning!c. Standardize blood choices based on maximum safety
3. Possible strategies (these are only estimates).
Blood needed in: Give:Less than 10 minutes(NOW!)
Uncrossmatched Group Oneg (may give O+ if maleor older female)
10-30 minutes Uncrossmatched ABOgroup and Rh type specific
Over 30 minutes Crossmatched ABO groupand Rh type specific
4. Documentationa. Eventually need signed physician statement on chart(not a waiver of Blood Bank responsibility)
b. Dont delay therapy to get signature! 5. Rh issues
a. All transfusion services should have a policy forstandard Rh types of blood to use in emergencies
b. For males and older females, it is acceptable to BEGIN by using O-positive blood in emergencies
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c. Use all possible means to give D-negative blood tochildbearing age and younger females
d. When D-neg blood is used, be prepared to switch to D- pos if massive use depletes inventory (see below)
B. Massive transfusion1. Several definitions
a. Transfusion of an amount of blood equal to the patients blood volume in 24 hours
b. Transfusion of 10+ units of blood in 24 hoursc. Transfusion of 50% of blood volume in 3 hoursd. Transfusion to replace blood loss of over 150 mL/min
2. Often follows emergency transfusion after trauma (seeabove), but also seen in:a. Gastrointestinal hemorrhage
b. Cardiac bypass or other cardiovascular surgeryc. Post-partum or other obstetrical hemorrhage
3. Complications largely from lethal triad that greatlyincreases mortality in massive transfusion settingsa. Coagulopathy
1) Thrombocytopenia due to platelet-poor transfusions2) Coagulopathy from dilution, factor-poor
transfusions; decreased body temp and pH alsoaffect coagulation
3) Many trauma patients have acute coagulopathy before arriving at hospital
4) See discussion of plasma transfusion in trauma inBBIII
5) If coagulopathic, usually have increased bleedingand have increased risk of shock and death
b. Acidosis1) pH declines in stored blood2) Citrate anticoagulant may also contribute to acidosis3) Tissue injury and poor perfusion leads to lactic
acidosisc. Hypothermia
1) Receiving large amounts of cold blood productsmay decrease body temperature
4. Summary of storage lesion of blood a. What goes down:
1) 2,3-DPG; see discussion below2) pH; see discussion below3) ATP; 50-60% levels at storage end4) Nitric oxide (NO); decreases oxygen delivery by
decreasing ability to vasodilate5) RBC membrane flexibility; makes it more difficult
to get into small capillaries b. What goes up:
1) Potassium; from hemolysis2) Free hemoglobin; from hemolysis
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b. Switching back1) Probably safest to wait until incompatible plasma
has been cleared (anti-A in above example)2) Use fresh sample to crossmatch vs. donor blood
8. Massive transfusion protocols (MTPs)a. Standardized, cookbook recipes for product use in
massive transfusion b. Most trauma centers have MTPs; technically required
by trauma certifying organizationsc. MTPs are designed to be proactive rather than reactived. Most MTPs today try to use 1:1 red cell to plasma
transfusion ratio, with platelets and cryo given variablydepending on the protocol
e. Studies are early but appear promising (problems: patient selection, randomization, confounding factors)
C. Organ transplantation1. 26,213 organs transplanted in US in 2010 (UNOS data)
a. 59% kidneys, 22% livers, 8% hearts b. Data relatively stable since 2001 or so
2. Transfusion needs greatest in heart and liver transplants,minimal in kidney transplants
3. General principles:a. ABO compatibility most important in organ
transplants (above HLA); in contrast to stem celltransplants.1) ABO-mismatched organs reserved for emergent,
dire circumstances; done more often now2) Plasma exchange and immunosuppression used to
minimize incompatible recipient ABO antibodies3) A 2 donor organs are special; low A antigen levels
on RBCs and endothelial cells may allow them togo to a group O or B recipient without a problem
b. Avoid HLA immunization before transplant if possible; especially in renal and cardiac transplant
c. Pre-renal transplant transfusion no longer requiredd. CMV prevention is only an issue when a seronegative
recipient gets an organ from a seronegative donor1) CMV seropositive donor is most likely source of
infection in seronegative recipients; no real need to provide CMV safe products
2) Most transplant centers are ok with leukocytereduced products for CMV risk reduction
4. Modifications to standard transfusions a. Leukocyte reduction: Standard (for HLA
immunization prevention and CMV-safe) b. Irradiation: Not standard (low risk of TA-GVHD), but
many centers do it routinely anywayc. ABO issues
1) Group A and O patients, use ABO-identical RBCs.
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2) Group AB and B may get A and O RBCs,respectively, to conserve more uncommon types.
3) In ABO-mismatched transplants, use type mostcompatible with donor and recipient for alltransfusions
4) The AB patient may also be a challenge with FFPtransfusion; group A FFP is probably OK duringsurgery
d. Rh issues1) Avoid D immunization before surgery if possible2) D-negative childbearing age females should get D-
negative blood unless supply cant support 3) In males and older females, may either give all D-
positive RBCs or use a staggered approach inhepatic and cardiac transplants (start with D-,switch to D+ in the middle, back to D- at the end)
5. Liver and heart transplantsa. Can be a HUGE stressor on transfusion services due to
often massive transfusion and coagulation issues1) Recent reports of nearly bloodless hepatic
transplantation surgeries have been seen2) Despite this, may severely strain supplies in many
cases b. Main issues are supply, communication and
responsiveness.1) Because of above, large amounts of all products
may be needed in a short time.2) Transplant team must notify Blood Bank ASAP
when decision to transplant is made.a) Transfusion services need this notice to increase
supply, test recipient for antibodies, findantigen-negative units if necessary
6. Kidney transplantsa. In the past, used multiple transfusions, but
erythropoietin therapy has decreased that need. b. Historical interest due to the discovery of transfusion-
associated immunosuppression in renal transplant patients transfused RBCs or WB before surgery1) Those transfused had less rejection.
c. Currently, main need is for CMV-safe products.5. Other organs
a. Most are similar in management to major surgery ofthe individual organs involved.
D. Platelet refractoriness1. Inability to respond to platelet transfusion with a
significant qualitative increase in platelet count.a. Big problem in multiply transfused patients
b. Nonimmune causes outweigh immune causes.
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1) Nonimmune: fever, splenomegaly, DIC, bleeding(?), drugs (eg, Amphotericin)
2) Immune: anti-HLA and/or anti-platelet antibodiesc. Management
1) Correct nonimmune causes, if possible.2) Consider fresher, ABO identical products.3) Check for immune causes (platelet antibody screen,
10 minute post count) if nonimmune not apparent.4) Strategies for platelet transfusion
a) HLA matchingi) Traditional strategy, using platelets from a
donor matching as many HLA class I (HLA-A and HLA-B) antigens as possible
ii) HLA matched usually means as close aswe could get.
iii) Match grades: A: 4 antigen match B1X: 1 antigen is cross-reactive B1U: 1 antigen is unknown B2UX: 1 unknown, 1 cross-reactive C: 3 antigen match D: 2 antigen match
iv) Irradiate platelets to prevent TA-GVHD. b) Platelet crossmatching
Patient serum vs. platelets in inventory (orsamples from potential donors); choose mostcompatible platelets
Uses solid-phase red cell adherence (SPRCA)technology (see BB1 section)
Some studies show more effectivenesscompared to HLA matching
c) Matching for antibody specificities (antibodyspecificity method) Determine specificity of antibodies, match for
platelets lacking those antibodies Analogous to giving someone with anti-K
units that are K negative Shown to be equal to HLA matching and
platelet crossmatching in responsed. Prevention is the best strategy; thus, leukocyte
reduction is used for patients likely to receive multipletransfusions (see BB3).E. Warm autoantibodies
1. Causes:a. Idiopathic (~ 50%)
b. Malignancies (CLL, NHL especially)c. Drugs (alpha-methyl dopa)d. Autoimmune disease (SLE)
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2) Some prefer to transfuse non-matched blood (or partially matched) and manage antibodies as theycome up.
b. Sickle-negative cells1) Many with sickle trait unaware of their condition.2) Some report ok results with sickle trait donors3) Best to ensure sickle-negative cells for transfusions.
G. Neonatal Transfusions (Birth to 4 months)1. Red blood cells:
a. Generally higher RBC transfusion thresholds than inadults; common thresholds follow:1) No symptoms: 8 g/dL2) Cardiopulmonary disease, major surgery: 10 g/dL3) Severe cardiopulmonary disease: 12-13 g/dL
b. Choice of anticoagulant-preservative is controversial1) Most adult RBC transfx are with AS-1, -3, or -52) Concern in neonates regarding mannitol (AS-1 and
-5) and excess volume3) Despite this, AS-RBCs are ok for small volume (10-
15 mL/Kg) RBC transfusions per studies4) For larger volumes, many use CPD or CPDA-1
units, or volume-reduce AS-RBCs beforetransfusion
c. Abbreviated pre-transfusion testing (see BBII, pg 14)2. Platelets
a. Also somewhat higher thresholds1) 100,000 for intracranial bleeds, ill premature babies2) 50,000 for other bleeding3) 20,000 for prophylaxis
b. Special concern: ABO compatibility1) Minor ABO mismatch that is ok in adult PLT
transfusions may be disastrous in neonates2) Remember incompatible plasma and smaller baby
volumes; keep ABO compatible if at all possible3. Plasma and Cryo
a. Similar indications to adults b. FP24 ok for use in neonates
4. Granulocytesa. Used often; may be more efficacious than in adults
b. Similar indications to adults, but may be given in babies that are less neutropenic (ANC
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b. Irradiation1) Some do routinely for all baby transfusions2) Probably only necessary in premature or those with
immunodeficienciesH. Therapeutic Apheresis (TA)
1. The use of apheresis to remove, replace, reduce, ormodify a pathologic cellular or plasma component
2. Guidelines from American Society for Apheresis (ASFA), published in Journal of Clinical Apheresis a. Category I: TA accepted, proven for primary therapy
b. Category II: TA accepted, useful for 2 nd-line therapyc. Category III: TA not proven but might be helpfuld. Category IV: TA not helpful and may be harmful
3. Types and methods available in U.S.:a. Therapeutic plasma exchange (TPE)
1) Remove pathologic plasma component and replacewith colloid (albumin, plasma) and/or crystalloids
2) Category I examples: TTP, Guillan-Barre, CIDP,Goodpasture syndrome, myasthenia gravis,cryoglobulinemia, antibody-mediated renal txrejection
b. Cytapheresis1) Removal of pathologic levels of native red cells,
platelets, or white cells (blasts)2) May include repl enishment with good cells (red
cell exchange in sickle cell disease, for example)3) Category I examples: Acute stroke in sickle cell
patients, severe babesiosis, leukostasis (blast crisis)a) Cat. II: Acute chest syndrome and stroke
prophylaxis in SCD b) Cat. II: Severe thrombocytosis in essential
thrombocythemiac. Extracorporeal photopheresis
1) Modification of patients mononuc. cells viaremoval, exposure to UV-A light in presence of
psoralen (crosslinks DNA), reinfusiona) Mechanism unclear; deactivates minority of cells
b) Side effects: Photosensitivity, hypotension2) Category I in cardiac allograft rejection, cutaneous
T-cell lymphoma/mycosis fungoides/Sezary3) Category II for skin chronic or acute GVHD
treatment in stem cell transplant patientsd. Selective adsorption (limited in US, broader Europe)
1) Selective removal of pathologic plasma components by various materials
2) Category I: LDL pheresis in familial hypercholest.3) Category II: Refractory ITP and RA
I. Hemolytic disease of the fetus/newborn (HDFN)1. Requirements
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a. The RAT model
1) R esponsea) Body responds to foreign antigen by making
antibody.2) Access
a) Antibody formed must be able to cross placenta. b) IgG1, IgG3, IgG4 cross, IgG2 and IgM do not.
3) T argeta) Target antigen must be present on fetal RBCs.
2. Manifestationsa. Severity depends on antibody involved.
1) Most severe: anti-D b. Lab findings
1) Anemia2) Indirect hyperbilirubinemia/jaundice3) Positive DAT (unless all cells destroyed)
c. Erythroblastosis fetalis 1) Abundant nucleated RBCs in circulation in response
to accelerated destructiond. Hydrops fetalis
1) Most severe form of HDFN, often fatal in utero2) EMH in liver may be so severe as to impair albumin
synthesis, leading to hydrops.3. ABO HDFN
a. Most common type of HDFN b. Generally mild or undiagnosed; Why?
1) Weak ABO antigen expression in utero2) Soluble plasma ABO antigens neutralize antibodies
c. Present in first (or any) pregnancyd. Group O moms, group A or B babiese. Lab testing
1) Cord blood DAT +/-a) Coated cells might be destroyed, or:
b) Cells may be very weakly coated2) Antibody screen neg. (screen cells are group O)3) Eluate from fetal cells + with A and/or B cells
f. Treatment1) No intervention is usually necessary, but if severe,
treat as outlined under Rh section below.4. Rh HDFN
a. Prototypical form of HDFN
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b. Not first pregnancy (unless mom was transfused)c. D-neg moms, D+ babies (ABO incompat reduces)d. Lab testing
1) Infant D+ (unl ess blocked D) a) Blocked D= false negative D antigen test due
to excessive coating of D+ cells with anti-D
2) DAT strongly positive3) Antibody screen: anti-D4) Eluate: anti-D5) Indirect bilirubin elevated6) Increased change in amniotic fluid optical density at
450 nm (Liley/Queenan graph)e. Treatment
1) Keep bilirubin < 20a) Lower threshold in premature babies
b) Indirect bilirubin can cross the blood/brain barrier and cause kernicterus (basal ganglia bilirubin deposition).
2) Phototherapy3) Exchange transfusion with D- blood as necessary
f. Prevention1) RhIG : commercially prepared anti-D
a) RhoGam, BayRho -D: IM forms b) WinRho, Rhophylac : IV or IM forms
2) Prevents D antigen recognition in vulnerable mom3) Obstetric indications
a) D-neg female at about 28 weeks gestation b) D-neg female < 72 hours of D+ infants birth c) D-neg female with pregnancy complications or
invasive procedures (amnio, cordocentesis)4) Contraindications
a) D-negative pregnant female who has alreadymade her own anti-D Think about prior RhIG, especially on exams
RhIG has a 25-day half-life, but may bedetectable for as long as six months
Usually distinguishable from maternal anti-D by weak reactions, IgM component, and lowtiter (almost always < 4)
b) D+ femalesc) D-neg mom, D-neg baby (no postpartum dose)
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5) Dosagea) One full dose vial (300 g or 1500 IU) per 30
ml of D+ whole blood (15 ml D+ RBCs) Mini -dose (50 g) sometimes used after 1 st
trimester abortion for < 5 ml whole blood b) Fetal blood screen (Rosette test)
Fetal D+ RBCs coated with anti-D thensurrounded by D+ indicator RBCs in rosettes
Qualitative (Yes/No) If positive, indicates need for quantitative test If negative, give 1 vial RhIG
c) Kleihauer-Betke Quantitative (but poorly reproducible) Acid-resistant Hgb F RBCs stain brightly on
blood smear Resul t is a percentage (% of moms blood
made of baby blood)d) Flow cytometry
Used in some centers rather than KB Samples many more cells and is considered
more accurate.e) Using quant. results (Tech Manual Method)
i) KB% x blood volume = baby blood in mom Use 5000 ml if no maternal weight given. If weight given, multiply weight x 70 ml/Kg
ii) Baby blood vol/30 = vials of RhIg needed Remember, one vial per 30 ml D+ blood
iii) Rounding rules If number after decimal is < 5, round up
If > 5, round up twice So, 3.4 would mean give 4 vials, while3.5 would mean to give 5 vials
iv) Example: D mom with D+ baby. KB=2% 0.02 x 5000 = 100 ml D+ baby blood 100/30 = 3.33 Round up once; give 4 vials
f) An easier and equally accurate way!i) KB% x 5/3 = number of vialsii) In above example, (2 x 5)/3 = 3.33iii) Note: 5 is moms BV in L ; use whatever
number is appropriateg. Compatibility testing for babies with Rh HDFN1) Moms serum m ay be used to crossmatch.
5. HDFN due to anti-Ka. Relatively common; particularly severe
b. Anti-K attaches to K antigen on early RBC precursorsand causes severe fetal anemia
c. Less hemolysis than other forms of HDFN (lesshyperbilirubinemia and reticulocytosis)
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II. Blood Bank CalculationsA. Chance of finding compatible units
1. Why bother?a. In real life: to allow Blood Bank techs to estimate the
difficulty in finding blood for patients with multiplealloantibodies
b. On exams: to torment you and make you feel generallyworthless and ignorant (dont let them succeed!) 2. Calculation
a. Find percentage of donors compatible:1) Likelihood of negativity = 1 Ag frequency2) Take likelihood of negativity for each antigen and
multiply3) Example:
a) Patient with anti-Jk a and anti-K (assume primarily Caucasian population)
b) Likelihood of Jk a negativity = 1 0.77 = 0.23
c) Likelihood of K negativity = 1 0.09 = 0.91d) Likelihood of Jk a and K negativity = 0.23 x 0.91= 0.2093, or 21% chance of this combinationgiven this donor pool
b. Estimate number of units you would have to test inorder to find x number of compatible units:1) Take number of units you need and divide by the
likelihood of finding antigen-negative blood.2) Example:
a) From above, looking for Jk a and K negativeunits, chance is 0.21.
b) For two units, divide 2 by 0.21 = 9.5 units thatyou would probably have to test to find two thatwere compatible (round to 10).
3. Pitfalla. Cant use for paired alleles (like Jk a and Jk b)
4. Chart for antigen frequencies (for reference; compiledfrom AABB Technical Manual ); see below
ANTIGEN CAUCASIANS AFRICANAMERICANS
D 85 92C 68 27c 80 96E 29 22e 98 98K 9 2k 99.8 99.9
Jk a 77 91Jk 72 43Fy a 66 10Fy 83 23
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M 78 70N 72 74S 55 31s 89 97
Le a 22 23Le 72 55
B. Corrected count increment (CCI)1. Why bother?
a. Objectively determine the response to platelettransfusion; corrects for size of patient & PLT dose
b. Most useful in very large or small recipients2. Calculation (most common formula)
dtransfuse plateletsof Number
10x)countPlatelet-count(PlateletBSA 11 pre post
BSA = body surface area in m 2 3. Interpretation
a. 7500 or above defines adequate response b. Two consecutive CCIs below 5000 defines
refractoriness.4. Pitfalls
a. Precount should be as near as possible to the time oftransfusion.
b. Post count is standardized for this equation at 1 hour post transfusion.
c. The 10 11 number is stated but is irrelevant (canceledout by the 10 11 from number of PLTs on bottom)
C. Cryo dosage for hypofibrinogenemia1. Why bother?
a. Again, often not done in real life (guesstimate) b. Calculates bags to raise level desired amount
2. Critical data neededa. Weight in Kg, hematocrit and recent fibrinogen level
3. Calculation1. Calculate Blood Volume
Body weight x 70 ml/Kg2. Calculate Plasma Volume (PV)
Blood volume x (1 - hematocrit)3. Calculate mg fibrinogen needed
Plasma volume x concentration change desired*Subtract desired level from current
(ie, 150 mg/dl 50 mg/dl)*Multiply level change by PV
(ie, 100 mg/dl x 3600 ml)*Divide the answer by 100 to correct fordifference in units (dl to ml)
4. Calculate bags of cryo needed Fibrinogen needed / 250 mg per bag
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III. Antibody IdentificationA. Whats, Whys, and Whens
1. What is a panel? a. RBCs from 8-20 different group O fully phenotyped
donors1) These RBCs come from a manufacturer, and the
antigens represented are regulated2) The presence or absence of each antigen is listed asa + or on each donors row of a sheet thataccompanies the RBCs
b. The panel RBCs are tested against serum or plasmafrom a patient or donor to determine incompatibility
2. Whya. Determine the target antigen of a potentially
significant non-ABO antibody b. Serves as a step to prevent a patient from receiving
incompatible RBCs
3. Whena. Evaluation of pretransfusion samples with apparentlysignificant antibodies on antibody screens
b. Evaluation of pregnant patients with positive screensc. Evaluation of blood donors with positive screensd. If testing suggests a NEW antibody in a patient with a
previously identified antibodyB. The process (see image below)
Check hi st or y Check autocont r ol Look at gener al pat t er n Look at whats N OT t her e (cr oss-outs) Look at what I S t her e Use special t echn iques as necessar y Ensure statistical signicance
1. Before you start:
a. Follow a defined process, and do so consistently
b. If on an exam, resist the temptation to skip to theanswers and try to fit an answer to the panel
c. Understand your technology (tube vs. non-tube,enhancement vs. non-enhancement, etc)
2. Check history a. Pregnancy, transfusions, medications, race, etc.
b. Impacts approximately 70% of cases3 Check autocontrol (AC)
a. Positive AC may alter the interpretation of the panel.
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a) Autoantibodies, alloantibodies to recentlytransfused cells, drug-related antibodies
b. Special techniques may be needed to understand the panel (autoadsorption, elution, etc.)
4. Look at general pattern for guidance a. Uniform reaction strength usually means a single
antibody b. Variable strength in multiple cells usually means
multiple antibodies (but may = dosage in one Ab)c. Variable reactions in multiple phases = multiple
antibodiesd. Take a glance at common antigens as a preview
5. Look at what is NOT there (cross-outs)
Fy a Fy b IS 37C IAT+ 0 0 0 0+ + 0 0 0
0 + 0 1+ 2+a. Exclude antibodies when the antigen is present but no
reaction is seen b. Start with cells that have totally negative reactions.
1) First two rows in above figure have no reaction.2) On line 1, since Fy a antigen is present, negative
reaction suggests that it is not antibody target.3) Indicate this by making a diagonal slash (cross-out)
through the Fy a at the top of the column. c. Only cross out double dose cells (except K )
1) Negatives may be nonpredictive when a single genecodes for the antigen (dosage)a) Some call this homozygous vs.
heterozygous (meaning, the donor in line 1 ishomozygous for the Fy a gene while the one inline 2 is heterozygous )
b) Double dose vs. single dose is betterterminology
2) So, on line 2, do not cross off either Fy a or Fy b.d. Horizontal vs vertical cross-outs
1) Common method: Do cross-outs all across a row,move down to the next negative row, do cross-outsall the way across that row, etc.
2) Alternative: Do cross-outs down a column, bloodgroup by blood group (often goes faster).
6. Look at what IS there a. Evaluate the cells that DO have reactionsb. Try to fit one antibody to all the reactions
1) If available, evaluate reactions as warm or cold . a) If all cold, look at Le a, Le b, M, N and P.
2) If a mix of warm and cold, skip to step c.
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b. Try to fit two or more antibodies in the same phase. 1) Phase only ap plicable in liquid tests2) If one antibody doesnt explain all reactions, try
two, then three, etc.c. Try to fit one warm and one cold antibody.
1) Only applicable if immediate spin results present2) If necessary, do cross-outs based on only warm,
then only cold reactions.7. Use special techniques as necessary
a. Patient phenotyping1) Helps confirm suspected specificity by showing
patient is NEGATIVE for the suspected antigentarget (as alloantibodies are vs. non-self antigens)
2) Shows possible antibodies in complicated workups b. Enzymes
1) Weaken or strengthen certain antigens2) Use to confirm preliminary conclusions
c. Adsorption1) Specific RBCs to remove antibody from serum.2) P atients own RBCs (autoadsorption) or RBCs of
known phenotype (alloadsorption).3) Remaining serum (a bsorbed) may be tested to
determine antibody specificity.d. Elution
1) Often used together with adsorption.2) Antibody rem oved (eluted) from RBC surface by
heat, cold, acid, solvent, or other treatment.3) A ntibody (eluate) may then be identified.
8. Ensure statistical significance 1) Two positive reactions when antigen is present and
two negatives when antigen is absent required byAABB reference lab standards
2) Labs may choose to require more or less (3 positives and 3 negatives commonly used)
9. Run screaming from the room1) We try to avoid getting to this stage!
B. Practical tips1. Time to burn?
a. Expect between 2 and 8 antibody panels on exams(occasionally, youll get lucky and get none) .
b. Dont spend too long on each one c. Moral: Guess and Go if necessary!
2. Anti-Da. Every time an anti-D shows up, your 1 st question in the
exam world should be: Has the patient had RhIG? b. RhIG can hang around and be detected for as long as
six months (half-life is about 25 days, though).3. Autoantibodies (see next section)4. Look for instant recognition (see next section)
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4. High-titer, low-avidity antibodies (HTLA) (panel D) a. Not a technically accepted term now, still mentioned
by blood bankers, though! b. Classically 1+ at AHG only, with negative autocontrol
1) Occasional positive autocontrol and DATsc. Still positive after many dilutions (high titer) but
weakly reacting (low avidity) d. Chido, Rodgers most common antigens
1) Complement components2) Neutralize with serum
e. Clinically insignificant (No HDFN, no HTRs)f. NOTE: This pattern could also be seen in other high-
frequency antibodies that may be significant!5. Reagent-related antibodies (panel E)
a. Antibodies against reagents used in testing (eg, preservatives in LISS)
b. Across-the-board positivity at IS/37 C, negative atIAT, positive autocontrol
c. DAT negative (due to washing step)d. Run reactions without offending reagent.
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IV. Sample antibody ID problems1. A 27-year- old female comes in for a tonsillectomy. Shes
never been transfused or pregnant. The above panel is performed. What is the antibody and what do you do?
*************************************************
2. A 19-year-old G3P1Ab1 female comes in at 28 weeks forroutine prenatal exam.
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page 22 Blood Bank Practical P}Chaffin (2/11/2013)
3. A 45-year-old male with a myelodysplastic syndrome andhistory of multiple transfusions.
4. A 69-year-old male with a chronic gastrointestinalhemorrhage and a hematocrit of 19%.
***************************************************5. A 41-year-old female with AML, transferred in from
another hospital; transfusion history unclear.
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P}Chaffin (2/11/2013) Blood Bank Practical page 23
Answers to Panels1. Anti-Le b
Single cold-reacting antibody. Anti-Le b reacting at thesetemperatures is not clinically significant, so no specificinterventions will be necessary. You might consider using
prewarmed crossmatches to eliminate the antibody
activity.
2. Anti-D Note that this is likely a gel or solid-phase panel (thoughit could be a liquid panel only recording the IATreactions). Single warm-reacting antibody. Fairly straight-forward identification. Check the clinical situation (anddont forget to ask about recent Rh IG injection orinfusion!).
3. Anti-K and anti-E
After your cross-offs, no single antibody explains all ofthe reactions, so you should try to fit two antibodies(again, this panel only shows IAT results, so no concernabout different phases here ). Anti-K and anti-E is the
best fit. Note the slightly weaker reactions in cell 6 due todosage.
4. Anti-Fy b and anti-Le a Just looking at this panel should make it very clear to youthat no single antibody will explain all of these results. Noantibody that I know of will give you this wide disparityin reaction strengths and temperatures. Postulate a singlewarm and a single cold antibody, and you come up with a
perfect fit with the above. However, if you also said thatyou cannot rule out anti-C, you are correct. In the realworld, you would have more work to do, but for our
purposes, you are done.
5. Anti-Jk a Did you notice the positive autocontrol? A positiveautocontrol with a mixed field pattern suggests anantibody against recently transfused antigens. This isclassic for Kidd antibodies. You should check the patientfirst to ensure that hes not in the midst of a delayedhemolytic reaction, and then check the prior hospital tosee if they picked up the anti-Jk a in their testing (itsentirely possible that it was not detectable).
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Pathology Review Cour seAdditional Resources
1. Booksa. AABB Technical Manual (17 th ed. is current)
b. AABB Standards (28 th ed. is current)c. Transfusion Medicine and Hemostasis, Chris Hillyer,
Beth Shaz, James Zimring, and Thomas Abshire, ed,;2009, Elsevier (WOW, what a great book! Everythingyou need in an easy-reading beauty of a spiral bound
book! Under 40 bucks on Amazon. Go get it, rightnow!)
d. Transfusion Medicine, 3 rd ed., Jeffrey McCullough,2012, McGraw-Hill (my former favorite; trulyexcellent while still being manageable. New edition
plagued by typos)e. Practical Transfusion Medicine, 2 nd ed., Murphy and
Pamphilon, 2005, Blackwell (fairly concise and practical).
f. Transfusion Medicine: Self-Assessment and Review,AABB Press (Review questions based topically onTech Manual).
2. Weba. Well, how about www.bbguy.org
b. Numerous presentations, articles, and lectures areavailable as well with a simple Google search