Review Cards - Immunohematology Flashcards
Criteria for whole blood donation - Allogenic
Age: >=16
HGB/HCT:
–Women: HGB >=12.5 g/dL OR HCT .+38%
–Men: HCG >13 g/dL OR HCT >39%
Temperature: <=37.5C (99.5F)
Venipuncture site: no infectious skin disease or scars indicative of drug use
Criteria for whole blood donation - Autologous
Age: as determined by medical director
HGB/HCT:
–HGB >=11.0 g/dL OR
–HCT >=33%
(NOTE: no donations within 72 hours of surgery)
Temperature: as determine by medical director; bacteremia is cause for deferral
Venipuncture site: as determined by medical director
Donor deferrals (AABB) - aspirin (donor sole source of platelets)
2 days
Donor deferrals (AABB) - measles (rubeola) vaccine
2 weeks
Donor deferrals (AABB) - mumps vaccine
2 weeks
Donor deferrals (AABB) - polio vaccine
2 weeks
Donor deferrals (AABB) - typhoid vaccine
2 weeks
Donor deferrals (AABB) - yellow fever vaccine
2 weeks
Donor deferrals (AABB) - rubella vaccine
4 weeks
Donor deferrals (AABB) - chicken pox (varicella-zoster) vaccine
4 weeks
Donor deferrals (AABB) - pregnancy
6 weeks
Donor deferrals (AABB) - whole blood donation
8 weeks
Donor deferrals (AABB) - after 2-unit RBC collection
16 weeks
Donor deferrals (AABB) - syphilis
12 months
Donor deferrals (AABB) - gonorrhea
12 months
Donor deferrals (AABB) - hepatitis B immune globulin (HBIG)
12 months
Donor deferrals (AABB) - mucous membrane exposure to blood
12 months
Donor deferrals (AABB) - skin penetration with sharp contaminated with blood or body fluids
12 months
Donor deferrals (AABB) - household or sexual contact with individual with hepatitis
12 months
Donor deferrals (AABB) - sexual contact with individual with HIV or at high risk
12 months
Donor deferrals (AABB) - incarceration in correctional facility for >72 consecutive hours
12 months
Donor deferrals (AABB) - travel to areas endemic for malaria
12 months
Donor deferrals (AABB) - recipient of blood, blood components, plasma-derived clotting factor concentrates, or transplant
12 months
Donor deferrals (AABB) - Babesia infection
at least 2 years
Donor deferrals (AABB) - malaria, or from an area endemic for malaria
3 years
Donor deferrals (AABB) - confirmed positive hepatitis B surface antigen (HBsAg)
permanent
Donor deferrals (AABB) - recipient of dura matter or pituitary growth hormone of human origin
permanent
Donor deferrals (AABB) - parenteral drug use
indefinite
Donor deferrals (AABB) - family history of Creutzfeldt-Jakob disease
indefinite
Donor deferrals (AABB) - repeated reactive test for anti-HBc on more than 1 occasion
indefinite
Donor deferrals (AABB) - positive hepatitis B virus (HBV) Nucleic Acid Amplification Testing (NAAT) result
indefinite
Donor deferrals (AABB) - repeatedly reactive test for anti-human T-lymphocyte virus (HTLV) on more than 1 occasion
indefinite
Donor deferrals (AABB) - present or past clinical evidence of infection with HIV, HCV, HTLV, or Trypanosoma cruzi
indefinite
Collection of whole blood - skin preparation
aseptic method, e.g., povidone-iodine scrub & prep solution
Collection of whole blood - volume of blood routinely collected
450 mL +/- 10% or 500 mL +/- 10%, depending on collection bag
Collection of whole blood - maximum volume
10.5 mL of blood per kg of donor’s weight, including samples for testing
Collection of whole blood - low-volume collections
-300-404 mL in 450 mL bag OR
-333-449 mL in 500-mL bag
-labeled “low volume”
-RBCs may be transfused, but plasma and platelets from “low volume” unit should be discarded
Collection of whole blood - volumes of anticoagulant
-63 mL anticoagulant for 450-mL collection
-70 mL for 500-mL collection
Collection of whole blood - time of collection
-usually <10 minutes
-if >15-20 minutes, unit may not be suitable for preparation of platelets or plasma
Collection of whole blood - samples for testing
from diversion pouch or by 2nd phlebotomy
Collection of whole blood - storage temperature of unit between collection & processing
20-24C if platelets are to be prepared; otherwise 1-6C
Apheresis - explanation
automated blood collection system that allows removal of 1 or more components of blood & return of remainder to donor
Apheresis - advantages
-allows collection of larger volumes of specific components
-can reduce number of donors to which patient is exposed
Apheresis - donor requirements
vary with procedure
Apheresis - components collected - RBCs
-2 units can be collected at same time from donors who are larger & have a higher HCT
-16 weeks between donations
Apheresis - components collected - Platelets
-plateletpheresis
-can collect HLA matched for patients who are refractory to random platelets
-can be leukoreduced during collection
-contain >=3 x 10^11 platelets
Apheresis - components collected - Plasma
plasmapheresis
Apheresis - components collected - Granulocytes
-leukapheresis
-not widely used to date
Apheresis - components collected - stem cells
-for bone marrow reconstitution in patients with cancer, leukemia, lymphoma
-autologous or HLA matched
Apheresis - therapeutic uses - therapeutic plasmapheresis
(plasma exchange) used to remove abnormal plasma proteins & replace with crystalloid, albumin, or FFP
Apheresis - therapeutic uses - therapeutic cytapheresis
used to remove cellular elements, such as an exchange transfusion for sickle cell patients, leukemic WBCs, lymphocytes (to induce immunosuppression)
Donor testing required by AABB and/or FDA - Typing
-ABO
-Rh (including weak D)
Donor testing required by AABB and/or FDA - antibody screen
antibody screen
Donor testing required by AABB and/or FDA - Syphilis testing
antibodies to Treponema pallidum or nontreponemal serological test for syphilis, e.g., RPR
Donor testing required by AABB and/or FDA - Hepatitis testing
-HBsAg
-Anti-HBc
-Anti-HCV
-HCV RNA (nucleic acid testing [NAT])
Donor testing required by AABB and/or FDA - HIV testing
-Anti-HIV-1/2
-HIV-1 RNA (NAT)
Donor testing required by AABB and/or FDA - other infectious disease testing
-Anti-HTLV-I/II
-West Nile virus RNA (NAT)
-Anti-Trypanosoma cruzi (FDA recommends 1-time donor screening)
-Zika virus (FDA recommends testing pooled donations)
Donor testing required by AABB and/or FDA - test to detect bacterial contamination of platelets
culture of platelets or FDA-approved rapid test (e.g., Pan Genera Detection [PGD] test)
Anticoagulant/Preservative Solutions - Acid citrate-dextrose (Formula A) - abbreviation
ACD-A
Anticoagulant/Preservative Solutions - Acid citrate-dextrose (Formula A) - RBC shelf life
21 days
Anticoagulant/Preservative Solutions - Acid citrate-dextrose (Formula A) - function
Citrate - prevents coagulation by chelating calcium
Dextrose - (glucose) supports ATP generation
Anticoagulant/Preservative Solutions - Acid citrate-dextrose (Formula A) - function
Citrate - prevents coagulation by chelating calcium
Dextrose - (glucose) supports ATP generation
Anticoagulant/Preservative Solutions - Acid citrate-dextrose (Formula A) - used for?
apheresis
Anticoagulant/Preservative Solutions - Citrate-phosphate-dextrose - abbreviation
CPD
Anticoagulant/Preservative Solutions - Citrate-phosphate-dextrose - RBC shelf life
21 days
Anticoagulant/Preservative Solutions - Citrate-phosphate-dextrose - function
-high pH preserves 2,3-DPG better
-better oxygen delivery
Anticoagulant/Preservative Solutions - Citrate-phosphate-dextrose-dextrose - abbreviation
CP2D
Anticoagulant/Preservative Solutions - Citrate-phosphate-dextrose-dextrose - RBC shelf life
21 days
Anticoagulant/Preservative Solutions - Citrate-phosphate-dextrose-dextrose - glucose
contains 100% more glucose than CPD
Anticoagulant/Preservative Solutions - Citrate-phosphate-dextrose with adenine - abbreviation
CPDA-1
Anticoagulant/Preservative Solutions - Citrate-phosphate-dextrose with adenine - RBC shelf life
35 days
Anticoagulant/Preservative Solutions - Citrate-phosphate-dextrose with adenine - function
-Adenine - increases ADP, which increases synthesis of ATP
-contains more glucose to sustain cells during longer storage
Additive solutions - purpose
extend shelf life of RBCs to 42 days
Additive solutions - constituents
-glucose for energy
-adenine to support ATP levels
Additive solutions - procedure
-plasma expressed from whole blood
100-110 mL additive transferred from attached satellite bag to RBCs within 72 hours of collection or per manufacturer’s instructions
Additive solutions - final hematocrit
55-65%
(HCT of RBCs without additive: 65-80%)
Additive solutions - examples
-Adsol (AS-1)
-Nutricel (AS-3)
-Optisol (AS-5)
Open System - explanation
-seal on unit is broken to attach external transfer bag
-exposure to air poses threat of bacterial contamination
Open system - effect on expiration date of component
-components stored at 1-6C must be used within 24 hours after system is open
-components stored at 20-24C must be used within 4 hours
Closed system -explanation
-sterility maintained though use of attached satellite bags or sterile connecting device that welds tubing from 1 bag to another
-no exposure to air
Closed system - effect on expiration date of component
no change
Blood components - RBCs - RBCs - preparation
-separated from whole blood by centrifugation or sedimentation any time before the expiration date of whole blood, OR
-collected by apheresis
Blood components - RBCs - RBCs - storage temperature
1-6*C
Blood components - RBCs - RBCs - shelf life
35 days in CPDA-1
Blood components - RBCs - RBCs - indications
inadequate tissue oxygenation
Blood components - RBCs - RBCs - HCT
should have HCT <=80%; otherwise not enough preservative to support RBCs
Blood components - RBCs - RBCs - HGB/HCT after transfusion of 1 unit
1 unit should increase HGB 1 g/dL or HCT 3%
Blood components - RBCs - RBCs adenine, saline added - preparation
additive solution added to RBCs following removal of most plasma
Blood components - RBCs - RBCs adenine, saline added - storage temperature
1-6*C
Blood components - RBCs - RBCs adenine, saline added - shelf life
42 days
Blood components - RBCs - RBCs adenine, saline added - indications
inadequate tissue oxygenation
What is the most commonly used RBC product?
RBCs adenine, saline added
Blood components - RBCs - RBCs frozen - preparation
-frozen in glycerol within 6 days of collection
-high glycerol (40%) method most commonly used
Blood components - RBCs - RBCs frozen - storage temperature
-frozen in high glycerol (40%): <=65C
-after removing glycerol (washing in decreased concentration of saline): 1-6C
Blood components - RBCs - RBCs frozen - shelf life
-frozen: 10 years
-after removing glycerol fix: 24 hours (unless a closed system used)
Blood components - RBCs - RBCs frozen - indications
inadequate tissue oxygenation
Blood components - RBCs - RBCs frozen - monitor glycerol removal
osmolality
Blood components - RBCs - RBCs frozen - what is removed?
all plasma, anticoagulant, WBCs, and platelets
Blood components - RBCs - RBCs frozen - safe for which patients?
IgA-deficient patients
Blood components - RBCs - RBCs frozen - used to?
store rare cells
Blood components - RBCs - Washed RBCs - preparation
RBCs washed with saline
Blood components - RBCs - Washed RBCs - storage temperature
1-6*C
Blood components - RBCs - Washed RBCs - shelf life
24 hours after washing
Blood components - RBCs - Washed RBCs - indications
history of severe allergic reaction (e.g., IgA, other plasma proteins)
Blood components - RBCs - Washed RBCs - % of RBCs lost in the process of washing?
about 20%
Blood components - RBCs - Washed RBCs - not a substitute for?
leukoreduced RBCs
Blood components - RBCs - Washed RBCs - not a substitute for?
leukoreduced RBCs
Blood components - RBCs - RBCs leukocytes reduced - preparation
filtration or apheresis processing
Blood components - RBCs - RBCs leukocytes reduced - storage temperature
1-6*C
Blood components - RBCs - RBCs leukocytes reduced - shelf life
-closed system: 35 days in CPDA-1
-open system: 24 hours
Blood components - RBCs - RBCs leukocytes reduced - shelf life
-closed system: 35 days in CPDA-1
-open system: 24 hours
Blood components - RBCs - RBCs leukocytes reduced - indications
history of febrile reaction
Blood components - RBCs - RBCs leukocytes reduced - must retain what % of original RBCs?
85%
Blood components - RBCs - RBCs leukocytes reduced - WBCs
<5 x 10^6
Blood components - RBCs - RBCs irradiated - preparation
irradiation at 2,500 cGy
Blood components - RBCs - RBCs irradiated - storage temperature
1-6*C
Blood components - RBCs - RBCs irradiated - shelf life
original outdate or 28 days from irradiation, whichever comes first
Blood components - RBCs - RBCs irradiated - indications
-immunodeficiency
-immunocompromised patients
-hematopoietic stem cell transplantation (HSCT) patients
-bone marrow transplant
-blood from blood relative
-intrauterine & neonatal transfusions
Blood components - RBCs - RBCs irradiated - used for prevention of?
graft-vs.-host disease
Blood components - RBCs - RBCs irradiated - gets rid of?
donor T cells
Blood components - Plasma & Derivatives - FFP - preparation
plasma separated from whole blood & frozen within 8 hours of collection
Blood components - Plasma & Derivatives - FFP - storage temperature
-frozen: <=-18C
-after thawing: 1-6C
Blood components - Plasma & Derivatives - FFP - shelf life
-frozen: 12 months
-after thawing: 24 hours
Blood components - Plasma & Derivatives - FFP - indications
deficiency of coagulation factors
Blood components - Plasma & Derivatives - FFP - contains?
all coagulation factors
Blood components - Plasma & Derivatives - FFP - thawed at?
30-37*C or by FDA-approved microwave
Blood components - Plasma & Derivatives - Cryoprecipitate - preparation
prepared by thawing FFP at 1-6*C, removing plasma, & re-freezing within 1 hour
Blood components - Plasma & Derivatives - Cryoprecipitate - storage temperature
Frozen: <=-18*C
After thawing: room temperature
Blood components - Plasma & Derivatives - Cryoprecipitate - shelf life
Frozen: 12 months
After thawing: single units 6 hours; pools 6 hours if sterile connecting device used; otherwise 4 hours
Blood components - Plasma & Derivatives - Cryoprecipitate - indications
fibrinogen & factor XIII deficiencies
Blood components - Plasma & Derivatives - Cryoprecipitate - diseases
used for hemophilia A and von Willebrand disease ONLY if factor VIII concentrate or recombinant factor preparations are not available
Blood components - Plasma & Derivatives - Cryoprecipitate - should contain?
> =80 IU of factor VIII and >=150 mg of fibrinogen
Blood components - Platelets - Platelets - preparation
-centrifugation of whole blood at RT within 8 hours of collection
-1st soft spin yields platelet-rich plasma
-2nd hard spin separates platelets from plasma
Blood components - Platelets - Platelets - storage temperature
20-24*C
Blood components - Platelets - Platelets - shelf life
-5 days from collection, with agitation
-after pooling: 4 hours
Blood components - Platelets - Platelets - indications
severe thrombocytopenia or abnormal platelet function
Blood components - Platelets - Platelets - requirements for unit
-40-70 mL plasma
->=5.5 x 10^10 platelets
-pH >=6.2
Blood components - Platelets - Platelets - 1 unit of platelets
should increase platelets by 5,000-10,000/uL in 75-kg patient
Blood components - Platelets - Platelets - # of units pooled
4-6
Blood components - Platelets - Apheresis platelets - preparation
apheresis
Blood components - Platelets - Apheresis platelets - storage temperature
20-24*C
Blood components - Platelets - Apheresis platelets - shelf life
5 days with agitation
Blood components - Platelets - Apheresis platelets - indications
severe thrombocytopenia or abnormal platelet function
Blood components - Platelets - Apheresis platelets - unit requirements
> =3.0 x 10^11 platelets (equivalent to 4-6 units)
Blood components - Platelets - Apheresis platelets - why used?
exposes recipient to fewer donors
Blood components - Platelets - Leukocyte-reduced platelets - preparation
WBCs removed by filtration or during apheresis processing
Blood components - Platelets - Leukocyte-reduced platelets - storage temperature
20-24*C
Blood components - Platelets - Leukocyte-reduced platelets - shelf life
open system: 4 hours
apheresis: 5 days
Blood components - Platelets - Leukocyte-reduced platelets - indications
-recurrent febrile reaction
-decrease risk of CMV transmission or HLA alloimmunization
Blood components - Platelets - Prestorage pooled platelets - preparation
4-6 ABO-identical platelets pooled using closed system
Blood components - Platelets - Prestorage pooled platelets - storage temperature
20-24*C
Blood components - Platelets - Prestorage pooled platelets - shelf life
5 days from collection
Blood components - Platelets - Prestorage pooled platelets - indications
-recurrent febrile reaction
-decrease risk of CMV transmission or HLA alloimmunization
Leukocyte Reduction (Leukoreduction) - purpose
to decrease WBCS to decrease febrile nonhemolytic transfusion reactions, transmission of CMV, & HLA alloimmunization
Leukocyte Reduction (Leukoreduction) - WBCs
<5 x 10^6
Leukocyte Reduction (Leukoreduction) - methods
-by apheresis processing
-by filtration during manufacture of components or after storage; prestorage leukocyte reduction is most effective; WBCs removed before they release cytokines
-use of filter during infusion
RBC storage lesion - increased
-lactic acid
-plasma K+
-plasma hemoglobin
-microaggregates
RBC storage lesion - decreased
-ATP
-2,3-diphosphoglycerate (2,3-DPG)
-pH
-glucose
-viable cells
-coagulation factors (the plasma has been removed)
RBC storage lesion - O2 dissociation curve
shift to left (increased HGB/O2 affinity, decreased O2 delivery to tissues)
Primary vs. Secondary response - Primary - stimulus
1st exposure to antigen
Primary vs. Secondary response - Primary - lag phase
days to months
Primary vs. Secondary response - Primary - type of antibody
IgM at first - may switch to IgG after 2-3 weeks (isotype switching)
Primary vs. Secondary response - Primary - titer
rises slowly, peaks, then declines
Primary vs. Secondary response - Secondary - stimulus
subsequent exposure to antigen
Primary vs. Secondary response - Secondary - lag phase
hours
Primary vs. Secondary response - Secondary - type of antibody
IgG
Primary vs. Secondary response - Secondary - titer
rises faster & higher, stays elevated longer
IgG Versus IgM - structure
IgG: monomer
IgM: pentamer
IgG Versus IgM - number of antigen binding sites
IgG: 2
IgM: 10
IgG Versus IgM - type of antibody
IgG: immune (adaptive)
IgM: naturally occurring
IgG Versus IgM - optimum temperature of reactivity
IgG: 37*C
IgM: 25*C or lower
IgG Versus IgM - reacts in saline?
IgG: no
IgM: yes
IgG Versus IgM - reacts best by indirect antiglobulin test?
IgG: yes
IgM: no
IgG Versus IgM - complement fixation
IgG: moderate
IgM: strong
IgG Versus IgM - causes transfusion reactions?
IgG: yes
IgM: not usually , except ABO
IgG Versus IgM - crosses placenta?
IgG: yes
IgM: no
IgG Versus IgM - causes HDFN?
IgG: yes
IgM: no
IgG Versus IgM - causes delayed hemolytic transfusion reactions?
IgG: yes
IgM: no
IgG Versus IgM - destroyed by sulfhydryl compounds (dithiothreitol [DTT], 2-mercaptoethanol [2-ME])?
IgG: no
IgM: yes
Factors that affect agglutination in tube testing - sensitization stage -
attachment of antibody to antigen
Factors that affect agglutination in tube testing - sensitization stage - temperature
clinically significant antibodies react best at 37*C
Factors that affect agglutination in tube testing - sensitization stage - pH
most antibodies react at pH 5.5-8.5
Factors that affect agglutination in tube testing - sensitization stage - ionic strength
reducing ionic strength of medium facilitates interaction of antibody with antigen (e.g., low ionic strength solution [LISS])
Factors that affect agglutination in tube testing - sensitization stage - antigen/antibody ratio
-too much antibody can cause prozone (false negative)
-optimum serum-to-cell ratio is 80:1
-usually 2 drops serum to 1 drop of 2-5% RBCs
Factors that affect agglutination in tube testing - sensitization stage - incubation time
-depends on medium
-usually 10-30 minutes
Factors that affect agglutination in tube testing - agglutination stage
formation of the lattice-type structure composed of the antigen-antibody bridges
Factors that affect agglutination in tube testing - agglutination stage - type of antibody molecule
IgM is larger, can span distance between RBCs more easily
Factors that affect agglutination in tube testing - agglutination stage - density of antigens & location on RBC surface
affects ease of attachment of antibodies
Factors that affect agglutination in tube testing - agglutination stage - zeta potential
-difference in charge between negatively-charged RBC surface & cloud of positive ions that surround RBCs
-reducing zeta potential allows RBCs to move closer together
-commercially available enhancement media reduces the zeta potential, allowing positively charged antibodies to get closer to negatively charged RBCs & increasing the RBC agglutination by IgG molecules
Comparison of tube, column, and solid-phase testing - tube testing - reaction container
glass test tubes
Comparison of tube, column, and solid-phase testing - tube testing - principle
-antibodies attach to corresponding antigens on RBCs, forming bridges between cells
-RBCS agglutinate
Comparison of tube, column, and solid-phase testing - tube testing - positive reaction
agglutinated RBCs or hemolysis
Comparison of tube, column, and solid-phase testing - tube testing - negative reaction
no agglutinated RBCs or hemolysis
Comparison of tube, column, and solid-phase testing - tube testing - adaptable to automation?
no
Comparison of tube, column, and solid-phase testing - column agglutination testing - reaction chamber
plastic microtube containing dextranacrylamide gel
Comparison of tube, column, and solid-phase testing - column agglutination testing - principle
-antigen-antibody reaction results in agglutinated RBCs
-gel acts as sieve
-large agglutinates can’t pass through, remain at top
-small agglutinates pass into gel
-unagglutinated cells go to bottom
Comparison of tube, column, and solid-phase testing - column agglutination testing - positive reaction
-agglutinated RBCs suspended in gel
-position indicates strength of reaction
-large agglutinates at top
Comparison of tube, column, and solid-phase testing - column agglutination testing - negative reaction
button of unagglutinated RBCs in bottom of microtube
Comparison of tube, column, and solid-phase testing - column agglutination testing - adaptable to automation?
yes
Comparison of tube, column, and solid-phase testing - solid-phase testing - reaction chamber
microplate with RBC membranes bound to surface of wells
Comparison of tube, column, and solid-phase testing - solid-phase testing - principle
-antibodies in sample attach to RBC antigens on surface of wells
-after incubation, unbound antibody removed by washing
-anti-IgG-labeled indicator RBCs added
-attach to antibodies bound to reagent RBC antigens during centrifugation
Comparison of tube, column, and solid-phase testing - solid-phase testing - positive reaction
indicator RBCs adhere diffusely to surface of well
Comparison of tube, column, and solid-phase testing - solid-phase testing - negative reaction
-no adherence of RBCs
-button of RBCs in bottom of well
Comparison of tube, column, and solid-phase testing - solid-phase testing - adaptable to automation?
yes
Comparison of tube, column, and solid-phase testing - solid-phase testing - advantages
-standardized
-more sensitive than tube testing
-reaction stable for 2 days
Comparison of tube, column, and solid-phase testing - tube testing - advantages
low cost
Comparison of tube, column, and solid-phase testing - column agglutination testing - advantages
-standardized
-more sensitive than tube testing
-reaction stable for 2-3 days; can be captured electronically
-antihuman globulin (AHG) tests don’t require washing or control cells
Grading reactions - tube versus column - 4+
Tube: one solid agglutinate
Column: solid band of agglutinated RBCs at top
Grading reactions - tube versus column - 3+
Tube: several large agglutinates
Column: band of agglutinated RBCs near top with a few staggered below
Grading reactions - tube versus column - 2+
Tube: medium-sized agglutinates, clear background
Column: agglutinates throughout
Grading reactions - tube versus column - 1+
Tube: small agglutinates, turbid background
Column: agglutinates predominantly in lower half of column with some RBCs at bottom
Grading reactions - tube versus column - mixed field
Tube: some agglutinated RBCs in sea of free RBCs
Column: layer of agglutinated RBCs at top & pellet of unagglutinated RBCs at bottom
Grading reactions - tube versus column - negative
Tube: no agglutinates
Column: well-defined pellet of unagglutinated RBCs at bottom
True or False. In tube testing, hemolysis is also a positive reaction.
True
ABO phenotype A - what is the genotype?
AA, AO
ABO phenotype B - what is the genotype?
BB, BO
ABO phenotype AB - what is the genotype?
AB
ABO phenotype O - what is the genotype?
OO
Frequency of ABO types - Type O
-European ancestry (%)
-African ancestry (%)
-Hispanic ancestry (%)
-Asian ancestry (%)
-European ancestry: 45%
-African ancestry: 49%
-Hispanic ancestry: 57%
-Asian ancestry: 40%
Frequency of ABO types - Type A
-European ancestry (%)
-African ancestry (%)
-Hispanic ancestry (%)
-Asian ancestry (%)
-European ancestry: 40%
-African ancestry: 27%
-Hispanic ancestry: 31%
-Asian ancestry: 27%
Frequency of ABO types - Type B
-European ancestry (%)
-African ancestry (%)
-Hispanic ancestry (%)
-Asian ancestry (%)
-European ancestry: 11%
-African ancestry: 19%
-Hispanic ancestry: 10%
-Asian ancestry: 25%
Frequency of ABO types - Type AB
-European ancestry (%)
-African ancestry (%)
-Hispanic ancestry (%)
-Asian ancestry (%)
-European ancestry: 4%
-African ancestry: 4%
-Hispanic ancestry: 2%
-Asian ancestry: 7%
ABO System - Group O
-antigen(s) on RBC?
-antibody(s) in serum?
-antigen(s) on RBC: neither
-antibody(s) in serum: anti-A, anti-B
ABO System - Group A
-antigen(s) on RBC?
-antibody(s) in serum?
-antigen(s) on RBC: A
-antibody(s) in serum: anti-B
ABO System - Group B
-antigen(s) on RBC?
-antibody(s) in serum?
-antigen(s) on RBC: B
-antibody(s) in serum: anti-A
ABO System - Group AB
-antigen(s) on RBC?
-antibody(s) in serum?
-antigen(s) on RBC: A and B
-antibody(s) in serum: none
Forward grouping:
Anti-A: 0
Anti-B: 0
Reverse grouping:
A cells: +
B cells: +
What is the blood type?
Type O
Forward grouping:
Anti-A: +
Anti-B: 0
Reverse grouping:
A cells: 0
B cells: +
What is the blood type?
Type A
Forward grouping:
Anti-A: 0
Anti-B: +
Reverse grouping:
A cells: +
B cells: 0
What is the blood type?
Type B
Forward grouping:
Anti-A: +
Anti-B: +
Reverse grouping:
A cells: 0
B cells: 0
What is the blood type?
Type AB
Anti-A: 0
Anti-B: 0
A1 cells: 0
B cells: 0
Possible cause?
Resolution?
Possible cause: missing isoagglutinins in group O
Resolution:
-incubate reverse grouping at RT for 30 minutes
-if still negative, incubate at 4*C for 15-30 minutes
-include controls: patient RBCs in 4% albumin, patient serum with O cells
Anti-A: 4+
Anti-B: 0
A1 cells: 1+
B cells: 4+
Possible cause?
Resolution?
Possible cause: A2 with anti-A1
Resolution:
-type RBCs with anti-A1 (Dolichos biflorus lectin)
-test serum with several additional A1, A2, & O cells
Anti-A: 4+
Anti-B: 4+
A1 cells: 2+
B cells: 2+
Possible cause?
Resolution?
Possible cause: Rouleaux
Resolution:
-use washed RBCs suspended in saline for forward grouping
-perform saline replacement technique in reverse grouping
Possible cause: AB with cold alloantibody
Resolution:
-perform antibody panel at RT
-if cold alloantibody identified, repeat reverse grouping with A1 & B cells that lack corresponding antigen
Anti-A: 3+
Anti-B: 4+
A1 cells: 1+
B cells: 0
Possible cause?
Resolution?
Possible cause: A2B with anti-A1
Resolution:
-type cells with anti-A1 (Dolichos biflorus lectin)
-test serum with additional A1, A2, & O cells
Anti-A: 4+
Anti-B: 2+
A1 cells: 0
B cells: 4+
Possible cause?
Resolution?
Possible cause: acquired B antigen
Resolution:
-check medical history for infection by GI bacteria (some have enzymes that convert A antigen to B-like antigen)
-retype RBCs with different monoclonal anti-B or acidified human anti-B (pH 6.0; doesn’t react with acquired B antigen)
Phenotype: Rh positive - genotype?
DD, Dd
Phenotype: Rh negative - genotype?
dd
Rh antigens - Fisher-Race: D
-Weiner:
-Rosenfield:
-Weiner: Rho
-Rosenfield: Rh1
Rh antigens - Fisher-Race: C
-Weiner:
-Rosenfield:
-Weiner: rh’
-Rosenfield: Rh2
Rh antigens - Fisher-Race: E
-Weiner:
-Rosenfield:
-Weiner: rh”
-Rosenfield: Rh3
Rh antigens - Fisher-Race: c
-Weiner:
-Rosenfield:
-Weiner: hr’
-Rosenfield: Rh4
Rh antigens - Fisher-Race: e
-Weiner:
-Rosenfield:
-Weiner: hr”
-Rosenfield: Rh5
Frequency of Rh antigens - D
-European ancestry:
-African ancestry:
-European ancestry: 85%
-African ancestry: 92%
Frequency of Rh antigens - C
-European ancestry:
-African ancestry:
-European ancestry: 68%
-African ancestry: 27%
Frequency of Rh antigens - E
-European ancestry:
-African ancestry:
-European ancestry: 29%
-African ancestry: 22%
Frequency of Rh antigens - c
-European ancestry:
-African ancestry:
-European ancestry: 80%
-African ancestry: 96%
Frequency of Rh antigens - e
-European ancestry:
-African ancestry:
-European ancestry: 98%
-African ancestry: 98%
Frequency of Rh genes - Rh^0
-antigens:
-European ancestry:
-African ancestry:
-Asian ancestry:
-antigens: Dce
-European ancestry: 4%
-African ancestry: 44%
-Asian ancestry: 3%
Frequency of Rh genes - Rh^1
-antigens:
-European ancestry:
-African ancestry:
-Asian ancestry:
-antigens: DCe
-European ancestry: 42%
-African ancestry: 17%
-Asian ancestry: 70%
Frequency of Rh genes - Rh^2
-antigens:
-European ancestry:
-African ancestry:
-Asian ancestry:
-antigens: DcE
-European ancestry: 14%
-African ancestry: 11%
-Asian ancestry: 21%
Frequency of Rh genes - Rh^z
-antigens:
-European ancestry:
-African ancestry:
-Asian ancestry:
-antigens: DCe
-European ancestry: <=0.01%
-African ancestry: <=0.01%
-Asian ancestry: 1%
Frequency of Rh genes - rh
-antigens:
-European ancestry:
-African ancestry:
-Asian ancestry:
-antigens: dce
-European ancestry: 37%
-African ancestry: 26%
-Asian ancestry: 3%
Frequency of Rh genes - rh’
-antigens:
-European ancestry:
-African ancestry:
-Asian ancestry:
-antigens: dCe
-European ancestry: 2%
-African ancestry: 2%
-Asian ancestry: 2%
Frequency of Rh genes - rh”
-antigens:
-European ancestry:
-African ancestry:
-Asian ancestry:
-antigens: dcE
-European ancestry: 1%
-African ancestry: <=0.01%
-Asian ancestry: <=0.01%
Frequency of Rh genes - rh^y
-antigens:
-European ancestry:
-African ancestry:
-Asian ancestry:
-antigens: dCE
-European ancestry: <=0.01%
-African ancestry: <=0.01%
-Asian ancestry: <=0.01%
Breaking the Rh Code - see r before h:
-think:
-example(s): rh’ = ?
-think: “big”
-rh’ = C
Breaking the Rh Code - see h before r:
-think:
-example(s): hr’ = ?
-think: “little”
-hr’ = c
Breaking the Rh Code - see R:
-think:
-example(s): Rh1 = ?
-think: presence of D
-Rh1 = DCe
Breaking the Rh Code - see r:
-think:
-example(s): rh’ = ?
-think: absence of D
-rh’ = dCe
Breaking the Rh Code - see 1 or ‘:
-think:
-example(s):
–Rh1 = ?
–rh’ - ?
–Rho = ?
-think: C
(if no 1 or ‘, then c)
-examples:
–Rh1 = DCe
–rh’ = dCe
-Rho = Dce
Breaking the Rh Code - see 2 or “:
-think:
-example(s):
–Rh2 = ?
–rh” = ?
–Rho = ?
-think: E
(if no 2 or “, then e)
-examples:
–Rh2 = DcE
–rh” = dcE
–Rho = Dce
Breaking the Rh Code - see 0:
-think:
-example(s): Rho = ?
-think: c + e
-Rho = Dce
Breaking the Rh Code - see Z or y:
-think:
-example(s): rh^y = ?
-think: C + E
-rh^y = dCE
Rh typing sera - High-protein anti-D - source
prepared from pools of human sera (immunized Rh-negative individuals)
Rh typing sera - High-protein anti-D - control
-same ingredients as reagent, except no anti-D
-should be purchased from same manufacturer as anti-D
Rh typing sera - High-protein anti-D - disadvantage
more false positives than low-protein reagents, e.g., RBCs with positive direct antiglobulin test (DAT)
Rh typing sera - Low-protein anti-D - source
mixture of monoclonal IgM & monoclonal or polyclonal IgG
Rh typing sera - Low-protein anti-D - control
-any negative typing reaction serves as control, e.g., negative reaction with anti-A or anti-B
-when RBCs react with all antisera (i.e., AB positive), run control recommended by manufacturer
-(usually patient RBCs with autologous serum or 6% or 8% albumin)
Rh typing sera - Low-protein anti-D - advantages
-most widely used
-lower rate of false positives with Ig-coated RBCs
Interpretation of Rh typing:
-anti-D: +
-control: 0
Rh type?
positive
Interpretation of Rh typing:
-anti-D: 0
-control: 0
Rh type?
negative*
*test for weak D if donor or infant of mother being evaluated for RhIG
Interpretation of Rh typing:
-anti-D: +
-control: +
Rh type?
invalid
List the causes of false positive Rh typing results.
-warm or cold autoagglutinins
-rouleaux
-polyagglutinable RBCs
-nonspecific agglutination due to ingredient or reagent, e.g., dye, preservative
-contaminated or incorrect reagent
List the causes of false negative Rh typing results.
-failure to add reagent
-RBC suspension too heavy
-resuspending cell button too vigorously
-contaminated or incorrect reagent
-blocking of antigen sites by antibodies, e.g., severe HDFN due to anti-D
Weak D testing - when performed?
when anti-D & Rh control are negative in Rh typing of donor or infant of mother being evaluated for RhIG
Weak D testing - reagent
not all anti-D reagents are appropriate for use
-refer to manufacturer’s package insert
Weak D testing - method
incubate Rh typing tubes at 37C for 15-60 minutes indirect antibody test (IAT)
*RBCs with positive DAT will react in any IAT test
Weak D testing - interpretation:
Anti-D = positive
Rh control = negative
Rh type?
Rh positive
Weak D testing - interpretation:
Anti-D = positive
Rh control = positive
Rh type?
invalid
Weak D testing - interpretation:
Anti-D = negative
Rh control = negative
Rh type?
Rh negative
Recipient type: Rh positive
Rh type patient can receive?
Rh positive OR Rh negative
Recipient type: weak D
Rh type patient can receive?
Rh positive OR Rh negative, depending on the weak D phenotype
Recipient type: Rh negative
Rh type patient can receive?
Rh negative only, especially women of childbearing age
(If Rh positive must be given in emergency, RhIG can be given to prevent immunization)
Frequency of other selected blood group antigens: K
-European ancestry:
-African ancestry:
-European ancestry: 9%
-African ancestry: 2%
Frequency of other selected blood group antigens: k
-European ancestry:
-African ancestry:
-European ancestry: 99.8%
-African ancestry: 100%
Frequency of other selected blood group antigens: Fya
-European ancestry:
-African ancestry:
-European ancestry: 68%
-African ancestry: 13%
Frequency of other selected blood group antigens: Fyb
-European ancestry:
-African ancestry:
-European ancestry: 80%
-African ancestry: 23%
Frequency of other selected blood group antigens: Jka
-European ancestry:
-African ancestry:
-European ancestry: 76%
-African ancestry: 92%
Frequency of other selected blood group antigens: Jkb
-European ancestry:
-African ancestry:
-European ancestry: 74%
-African ancestry: 48%
Frequency of other selected blood group antigens: M
-European ancestry:
-African ancestry:
-European ancestry: 79%
-African ancestry: 74%
Frequency of other selected blood group antigens: N
-European ancestry:
-African ancestry:
-European ancestry: 70%
-African ancestry: 75%
Frequency of other selected blood group antigens: S
-European ancestry:
-African ancestry:
-European ancestry: 55%
-African ancestry: 30%
Frequency of other selected blood group antigens: s
-European ancestry:
-African ancestry:
-European ancestry: 90%
-African ancestry: 92%
Frequency of other selected blood group antigens: Lea
-European ancestry:
-African ancestry:
-European ancestry: 22%
-African ancestry: 23%
Frequency of other selected blood group antigens: Leb
-European ancestry:
-African ancestry:
-European ancestry: 72%
-African ancestry: 55%
Frequency of other selected blood group antigens: P1
-European ancestry:
-African ancestry:
-European ancestry: 79%
-African ancestry: 94%
I system - I antigen
Adult cells:
Cord cells:
i adult cells:
Adult cells: much
Cord cells: trace
i adult cells: trace
I system - i antigen
Adult cells:
Cord cells:
i adult cells:
Adult cells: trace
Cord cells: much
i adult cells: much
Antibody characteristics - naturally occurring
-ABO
-Lewis
-P1
-MN
-Lu^a
Antibody characteristics - clinically significant
-ABO
-Rh
-Kell
-Duffy
-SsU
Antibody characteristics - warm antibodies
-Rh
-Kell
-Duffy
-Kidd
Antibody characteristics - cold antibodies
-M
-N
-P1
Antibody characteristics - usually only react in AHG
-Kell
-Duffy
-Kidd
Antibody characteristics - can react in any phase of testing
Lewis
Antibody characteristics - detection enhanced by enzyme treatment of test cells
-Rh
-Lewis
-Kidd
-P1
Antibody characteristics - not detected with enzyme treatment of test cells
-M
-N
-Duffy
Antibody characteristics - enhanced by acidification
M
Antibody characteristics - show dosage
-Rh other than D
-MNS
-Duffy
-Kidd
Antibody characteristics - bind complement
-I
-Kidd
-Lewis
Antibody characteristics - cause in vitro hemolysis
-ABO
-Lewis
-Kidd
-Vell
-some P1
Antibody characteristics - labile in vivo & in vitro
Kidd
Antibody characteristics - common cause of anamnestic response (delayed transfusion reaction)
Kidd
Antibody characteristics - associated with paroxysmal nocturnal hemoglobinuria
Anti-P
Antibody characteristics - associated with cold agglutinin disease & Mycoplasma pneumoniae infections
Anti-I
Antibody characteristics - associated with infectious mononucleosis
anti-i
Antigen-Antibody enhancement - reagent - albumin
22% bovine serum albumin
-reduces net negative charge of RBCs, allowing them to come closer together
Antigen-Antibody enhancement - reagent - LISS
-reduces ionic strength of suspending medium (lowers zeta potential), allowing antigens & antibodies to move closer together more rapidly
-reduces incubation time for IAT
Antigen-Antibody enhancement - reagent - Polyethylene glycol (PEG)
-increases antibody uptake
-used for detection & ID of weak IgG antibodies
Antigen-Antibody enhancement - reagent - enzymes
-ficin & papain most commonly used
-reduce RBC surface charge by cleaving sialic acid molecules
-M, N, S, Fya, Fyb antigens destroyed
Antihuman Globulin (AHG) serum - type - Polyspecific (broad spectrum) - detects
IgG & C3d
Antihuman Globulin (AHG) serum - type - Polyspecific (broad spectrum) - used for?
DAT
Antihuman Globulin (AHG) serum - type - Polyspecific (broad spectrum) - DAT positive
monospecific sera used for follow-up testing
Antihuman Globulin (AHG) serum - type - Polyspecific (broad spectrum) - advantage
may detect complement-binding antibody more readily
Antihuman Globulin (AHG) serum - type - Polyspecific (broad spectrum) - disadvantage
reaction due to complement binding by clinically insignificant cold antibody
Antihuman Globulin (AHG) serum - type - Monospecific - Anti-IgG - detects
IgG
Antihuman Globulin (AHG) serum - type - Monospecific - Anti-IgG - used for?
routine compatibility tests & antibody ID
Antihuman Globulin (AHG) serum - type - Monospecific - Anti-IgG - advantage
detects clinically significant antibodies
Antihuman Globulin (AHG) serum - type - Monospecific - Anti-C3d or anti-C3b-C3d - detects
complement
Antihuman Globulin (AHG) serum - type - Monospecific - Anti-C3d or anti-C3b-C3d - detects
complement
Antihuman Globulin (AHG) serum - type - Monospecific - Anti-C3d or anti-C3b-C3d - helpful in investigation of what?
immune hemolytic anemia
Antiglobulin testing - Direct (DAT) - detects
in vivo sensitization of RBCs by IgG antibody
Antiglobulin testing - Direct (DAT) - specimen
EDTA red cells preferred
Antiglobulin testing - Direct (DAT) - incubation
none required
Antiglobulin testing - Direct (DAT) - application
-HDFN
-transfusion reaction
-autoimmune hemolytic anemia
-drug-induced hemolytic anemia
Antiglobulin testing - Direct (DAT) - false positives
-complement binding in vitro if RBCs are taken from red-top tube & broad-spectrum AHG used
-septicemia
-contamination of specimen
-Wharton jelly in cord blood
-overcentrifugation
Antiglobulin testing - Direct (DAT) - false negatives
-interruption in testing
-contamination, improper storage
-failure to add AHG
-neutralization of AHG from inadequate washing
-dilution of AHG by residual saline
-undercentrifugation
Antiglobulin testing - Indirect (IAT) - detects
in vitro sensitization of RBCs by IgG antibody
Antiglobulin testing - Indirect (IAT) - specimen
serum, plasma, RBCs
Antiglobulin testing - Indirect (IAT) - incubation
-serum or plasma with reagent RBCs OR
-RBCs with reagent antiserum
Antiglobulin testing - Indirect (IAT) - application
-antibody screen
-crossmatch
-RBC phenotyping
-weak D testing
-antibody ID
Antiglobulin testing - Indirect (IAT) - false positives
-cells with positive DAT
-overcentrifugation
Antiglobulin testing - Indirect (IAT) - false negatives
-interruption in testing
-contamination, improper storage
-failure to add AHG
-neutralization of AHG from inadequate washing
-dilution of AHG by residual saline
-undercentrifugation
-underincubation
Antibody identification - reactions: same strength & in 1 phase only - possible explanation?
suggestive of single antibody
Antibody identification - reactions: varying strength - possible explanation?
-multiple antibodies
-antigens exhibiting dosage
Antibody identification - reactions: in different phases - possible explanation?
-combination of warm & cold antibodies
-antibody with wide thermal range
Antibody identification - reactions: all cells in AGH, autocontrol negative - possible explanation?
-multiple antibodies
-antibody to high frequency antigen
Antibody identification - reactions: all cells at 37*C, negative in AHG, autocontrol positive - possible explanation?
rouleaux
Antibody identification - reactions: all cells in AHG, autocontrol positive - possible explanation?
warm autoantibody
Interpreting antibody panels - rule out (cross out)
- On row of antigens at top of antigram, cross out those that are present on cells that didn’t react in any phase.
- Circle antigens that aren’t crossed out. An antibody to 1 or more of these may be present.
- Look for matching patterns, e.g., if serum only reacted with cells #3 & #5, and E antigen is only on cells #3 and #5, the antibody is most likely anti-E
An antibody will react with all cells that possess the corresponding antigen (except for weak antibodies that only react with homozygous cells). An antibody won’t react with any cells that lack the corresponding antigen.
Interpreting antibody panels - testing with selected cells
If other antibodies can’t be ruled out, further testing with selected cells might be required. Cells selected for testing should be positive for only 1 antigen corresponding to antibodies in question, e.g., if pattern reactivity matches anti-Jka, but anti-K and anti-S can’t be ruled out, serum should be tested with cells of the following type:
*Jk(a-), K+, S-
*Jk(a-), K-, S+
*Jk(a+), K-, S-
Homozygous cells are preferred to avoid missing weak antibodies that demonstrate dosage.
Interpreting antibody panels - Confirmation
Conclusive ID requires testing sufficient numbers of cells that are positive and negative for the corresponding antigen, e.g., 3 positive & 3 negative. Some labs use other combinations that provide the same probability (p) value of <=0.05 (I.e., 95% confidence level that the antibody ID is correct). Once the antibody is identified, type patients RBCs for corresponding antigen. Should be negative.
Cold antibodies - anti-A1
-only found in subgroups of A
-agglutinates A1 and A1B cells, but not A2 or O.
Cold antibodies - anti-I
-agglutinates all adult cells, except i adult
-doesn’t agglutinate cord cells
Cold antibodies - anti-i
agglutinates cord cells more strongly than adult cells
Cold antibodies - anti-H
-most common in A1 and A1B
-agglutinates O cells most strongly, followed by A2 and B; least likely in A1B
Cold antibodies - anti-IH
-most common in A1 and A1B
-agglutinates cells that possess both I and H
-agglutinates adult O cells most strongly
-weaker reaction with A1 cells
-doesn’t agglutinate cord cells
Compatibility testing - specimen collected within how many days of transfusion if patient has been pregnant or transfused in preceding 3 months?
3 days
Compatibility testing - should the Rh type on the donor unit be repeated if it is labeled Rh neg?
yes
Compatibility testing - should weak D be performed on the donor unit if it is labeled Rh neg?
no
Compatibility testing - how many determinations of the recipient’s ABO group per AAB Standard 5.14.5?
2
Compatibility testing - what samples should the crossmatch be performed on?
recipient serum & donor RBCs
Compatibility testing - how long should the patient specimen & unit segment be retained for after transfusion?
7 days (kept at 1-6*C)
Crossmatches - antiglobulin - explanation
recipient serum + donor RBCs tested through AHG
Crossmatches - antiglobulin - when performed?
if recipient has or had clinically significant antibody
Crossmatches - immediate spin - explanation
recipient serum + donor RBCs tested in IS only
Crossmatches - immediate spin - when performed
if recipient doesn’t have & never had clinically significant antibody
Crossmatches - immediate spin - detects?
ABO incompatibility
Crossmatches - electronic - explanation
computer check of donor ABO & Rh type AND recipient ABO & Rh type
Crossmatches - electronic - when performed
if recipient doesn’t have & never had clinically significant antibody
Crossmatches - electronic - detects?
ABO incompatibility
Crossmatches - electronic - ABO typing of recipient must be done how many times?
twice
The Major Crossmatch - will?
-detect ABO incompatibility
-detect most antibodies against donor cells
The Major Crossmatch - will NOT?
-detect all ABO typing errors
-detect most Rh typing errors
-detect all antibodies
-detect platelet or leukocyte antibodies
-prevent immunization
-ensure normal survival of RBCs
Incompatible Crossmatches: Negative antibody screen, incompatible IS crossmatch.
-possible cause?
-resolution?
Possible cause: ABO incompatibility
Resolution: retype donor & recipient; crossmatch with ABO-compatible donor
Incompatible Crossmatches: 1 antibody screening cell & 1 donor positive in AHG
-possible cause?
-resolution?
Possible cause: Alloantibody
Resolution: ID the antibody; crossmatch units negative for corresponding antigen
Incompatible Crossmatches: Antibody screening cells & all donor except 1 negative at 37*C & in AHG; 1 donor positive in AHG only
-possible cause?
-resolution?
Possible cause: positive DAT on donor
Resolution: perform DAT on unit; if positive, return to collecting facility
Incompatible Crossmatches: Antibody screening cells, donors, & autocontrol positive in AHG
-possible cause?
-resolution?
Possible cause: warm autoantibody
Resolution: important to detect any underlying clinically significant alloantibodies prior to transfusion; Adsorption testing is typically required; if patient situation is life-threatening, consult with medical director and provide transfusion.
Incompatible Crossmatches: Antibody screening cells, donors, & autocontrol positive at IS & 37*C, but negative in AHG
-possible cause?
-resolution?
Possible cause: rouleaux
Resolution: saline replacement technique
Transfusion of Non-Group-Specific RBCs:
-Patient type: O
-RBC type patient can receive?
-Plasma type patient can receive?
RBC type patient can receive: O only
Plasma type patient can receive: O, A, B, AB
Transfusion of Non-Group-Specific RBCs:
-Patient type: A
-RBC type patient can receive?
-Plasma type patient can receive?
RBC type patient can receive: A, O
Plasma type patient can receive: A, AB
Transfusion of Non-Group-Specific RBCs:
-Patient type: B
-RBC type patient can receive?
-Plasma type patient can receive?
RBC type patient can receive: B, O
Plasma type patient can receive: B, AB
Transfusion of Non-Group-Specific RBCs:
-Patient type: AB
-RBC type patient can receive?
-Plasma type patient can receive?
RBC type patient can receive: AB, A, B, O
Plasma type patient can receive: AB
Pretransfusion testing - RBCs - crossmatch?
yes
-can be electronic crossmatch if negative antibody screen & no record of previously detected antibodies
Pretransfusion testing - Plasma - crossmatch?
no
-must be ABO compatible with recipient RBCs
Pretransfusion testing - Cryoprecipitate - crossmatch?
no
-all ABO groups acceptable; Rh type not considered
Pretransfusion testing - Platelets - crossmatch?
No*
*for apheresis platelets containing >2 mL RBCs, ABO compatibility & crossmatch required
-any ABO group acceptable; compatible with recipient RBCs recommended; ABO-identical preferred
List the conditions for reissue of RBCs.
-maintained at 1-10*C
-closure wasn’t broken
-at least 1 sealed segment remains attached to unit
-unit is inspected before release
-records indicate that blood has been inspected & is acceptable for reissue
List the emergency transfusion thresholds.
-if time allows for typing, give ABO & Rh compatible; otherwise give O-neg RBCs
-label must indicate that crossmatch wasn’t completed
-physician must sign emergency release
-routine testing must be completed & physician notified immediately of any incompatibility
Transfusion-associated infections - HIV - prevention
donor testing
Transfusion-associated infections - HIV - estimated risk
1 in 1,467,000 transfusions
Transfusion-associated infections - HIV - window period with NAT
9 days
Transfusion-associated infections - Hepatitis B -prevention
donor testing
Transfusion-associated infections - Hepatitis B - estimated risk
1 in 280,000 transfusions
Transfusion-associated infections - Hepatitis C - prevention
donor testing
Transfusion-associated infections - Hepatitis C - estimated risk
1 in 1,149,000 transfusions
Transfusion-associated infections - Hepatitis C - window period with NAT
7.4 days
Transfusion-associated infections - HTLV-II and -II - prevention
donor testing
Transfusion-associated infections - HTLV-II and -II - estimated risk
rare in the US
Transfusion-associated infections - Syphilis - prevention
donor testing
Transfusion-associated infections - Syphilis - risk
-limited risk in refrigerated or frozen components - spirochetes cannot survive the cold
-HIGHEST risk from platelets
Transfusion-associated infections - Malaria - prevention
donor testing
Transfusion-associated infections - Malaria - transmission
Plasmodium is transmitted in RBCs
Transfusion-associated infections - Babesiosis - prevention
donor testing
Transfusion-associated infections - Babesiosis - transmission
B. microti is transmitted in RBCs
Transfusion-associated infections - Chagas disease - prevention
donor testing
Transfusion-associated infections - Chagas disease - transmission
Trypanosoma cruzi is transmitted in blood.
-potential risk in donors from Central & South America
Transfusion-associated infections - CMV - prevention
-selected donor testing (not routine)
-use of CMV-neg or leukoreduced components for patients at risk
Transfusion-associated infections - CMV - transmission
CMV transmitted in WBCs
Transfusion-associated infections - CMV - risk
risk to CMV-neg immunocompromised patients & premature infants of CMV-neg mothers
Transfusion-associated infections - Zika virus - prevention
donor testing
Transfusion-associated infections - Zika virus - risk
-no confirmed transfusion-transmission cases of Zika in the US, but Brazil has reported platelet transfusion transmission
Transfusion-associated infections - West Nile virus - prevention
donor testing
Transfusion-associated infections - West Nile virus - NAT
used to screen all prospective donors in the US - most people are unaware they are infected
Transfusion-associated infections - Sepsis - prevention
-donor screening (history, temp)
-aseptic technique
-use of diversion pouches (1st 30-45 mL of blood goes into pouch so that skin plug doesn’t enter unit)
-tests to detect bacterial contamination of platelets (e.g., culturing 24 hours after collection)
Transfusion-associated infections - Sepsis - highest risk from what blood component?
platelets - due to RT storage
Acute immunologic transfusion reactions - Hemolytic, intravascular - clinical signs
-fever
-chills
-shock
-renal failure
-DIC
-pain in chest, back or flank
Acute immunologic transfusion reactions - Hemolytic, intravascular - cause
immediate destruction of donor RBCS by recipient antibody
Acute immunologic transfusion reactions - Hemolytic, intravascular - laboratory findings
-post-transfusion specimen:
–HGB in urine & serum
–mixed field DAT (unless donor cells are all destroyed)
–decreased haptoglobin, HGB, & HCT
Acute immunologic transfusion reactions - Hemolytic, intravascular - usually due to?
transfusion of ABO-incompatible blood
Acute immunologic transfusion reactions - Febrile - clinical signs
temperature increased >=1C or 2C during or shortly after transfusion, with no other explanation
Acute immunologic transfusion reactions - Febrile - cause
anti-leukocyte antibodies or cytokines
Acute immunologic transfusion reactions - Febrile - laboratory findings
none
Acute immunologic transfusion reactions - Febrile - most often in?
multiply transfused patients or women with multiple pregnancies
Acute immunologic transfusion reactions - Febrile - what type of unit should be given if a patient has had a febrile reaction before?
leukoreduced components
Acute immunologic transfusion reactions - Febrile - premedicate?
antipyretics (aspirin, acetominophen)
Acute immunologic transfusion reactions - Allergic - clinical signs
-hives (urticaria)
-wheezing
Acute immunologic transfusion reactions - Allergic - cause
foreign plasma proteins
Acute immunologic transfusion reactions - Allergic - laboratory findings
none
Acute immunologic transfusion reactions - Allergic - treatment?
antihistamines
Acute immunologic transfusion reactions - Anaphylactic - clinical signs
-pulmonary edema
-bronchospasms
Acute immunologic transfusion reactions - Anaphylactic - cause
anti-IgA in IgA-deficient recipient
Acute immunologic transfusion reactions - Anaphylactic - laboratory findings
none
Acute immunologic transfusion reactions - Anaphylactic - treatment
epinephrine
Acute immunologic transfusion reactions - Anaphylactic - transfuse with?
washed cellular blood products
Acute immunologic transfusion reactions - Transfusion-related acute lung injury (TRALI) - clinical signs
-fever
-chills
-coughing
-respiratory distress
-fluid in lungs
-decreased blood pressure within 6 hours of transfusion
LIFE THREATENING
Acute immunologic transfusion reactions - Transfusion-related acute lung injury (TRALI) - cause
-unknown
-possibly donor antibodies to WBC antigens
Acute immunologic transfusion reactions - Transfusion-related acute lung injury (TRALI) - laboratory findings
none
Acute immunologic transfusion reactions - Transfusion-related acute lung injury (TRALI) - related to?
infusion of plasma
-all components have been implicated
Acute immunologic transfusion reactions - Transfusion-related acute lung injury (TRALI) - reduction of what appears to be reducing TRALI fatalities?
reduced use of plasma from female donors
Acute nonimmunologic transfusion reactions - Sepsis - clinical signs
-fever
-chills
-decreased BP
-cramps
-diarrhea
-vomiting
-muscle pain
-DIC
-shock
-renal failure
Acute nonimmunologic transfusion reactions - Sepsis - cause
bacterial contamination
Acute nonimmunologic transfusion reactions - Sepsis - laboratory findings
-positive gram stain & culture on unit
-positive blood culture on patient
Acute nonimmunologic transfusion reactions - Transfusion-associated circulatory overload (TACO) - clinical signs
-coughing
-cyanosis
-difficulty breathing
-pulmonary edema
Acute nonimmunologic transfusion reactions - Transfusion-associated circulatory overload (TACO) - cause
too large a volume or too rapid rate of infusion
Acute nonimmunologic transfusion reactions - Transfusion-associated circulatory overload (TACO) - laboratory findings
none
Acute nonimmunologic transfusion reactions - Transfusion-associated circulatory overload (TACO) - occurs most often in what patients?
-children
-cardiac & pulmonary patients
-elderly
-patients with chronic anemia
Acute nonimmunologic transfusion reactions - Nonimmune hemolysis - clinical signs
variable
Acute nonimmunologic transfusion reactions - Nonimmune hemolysis - cause
destruction of RBCs due to extremes of temperature, addition of meds to unit
Acute nonimmunologic transfusion reactions - Nonimmune hemolysis - laboratory findings
-hemoglobinuria
-hemoglobinemia
Acute nonimmunologic transfusion reactions - Hypothermia - clinical signs
cardiac arrhythmia
Acute nonimmunologic transfusion reactions - Hypothermia - cause
rapid infusion of large amounts of cold blood
Acute nonimmunologic transfusion reactions - Hypothermia - laboratory findings
none
Acute nonimmunologic transfusion reactions - Hypothermia - prevention
use blood warmer for rapid infusions
Delayed transfusion reactions - Immunologic - hemolytic, extravascular - clinical signs
-fever
-anemia
-mild jaundice
-2-14 days after transfusion
Delayed transfusion reactions - Immunologic - hemolytic, extravascular - cause
donor RBCs sensitized by recipient IgG antibody & removed from circulation
Delayed transfusion reactions - Immunologic - hemolytic, extravascular - laboratory findings
-increased bilirubin
-mixed-field DAT
-decreased haptoglobin
-decreased HGB & HCT
-positive antibody screen
Delayed transfusion reactions - Immunologic - hemolytic, extravascular - may be due to?
anamnestic response
Delayed transfusion reactions - Immunologic - hemolytic, extravascular - which antibodies are the most common cause?
Kidd
Delayed transfusion reactions - Immunologic - hemolytic, extravascular - life threatening?
usually NO
Delayed transfusion reactions - Immunologic - alloimmunization - clinical signs
none, unless subsequently exposed to same foreign antigens
Delayed transfusion reactions - Immunologic - alloimmunization - cause
development of antibodies to foreign RBCs, WBCs, platelets, plasma proteins following transfusion
Delayed transfusion reactions - Immunologic - alloimmunization - laboratory findings
-antibody to RBCs detected in antibody screen
-others require specialized testing
Delayed transfusion reactions - Immunologic - alloimmunization - components that should be used for patients with WBC antibodies?
leukoreduced components
Delayed transfusion reactions - Immunologic - Transfusion-associated graft-vs.-host disease (TA-GVHD) - clinical signs
-rash
-nausea
-vomiting
-diarrhea
-fever
-pancytopenia
USUALLY FATAL
Delayed transfusion reactions - Immunologic - Transfusion-associated graft-vs.-host disease (TA-GVHD) - cause
viable T lymphocytes in donor blood attack recipient
Delayed transfusion reactions - Immunologic - Transfusion-associated graft-vs.-host disease (TA-GVHD) - laboratory findings
none
Delayed transfusion reactions - Immunologic - Transfusion-associated graft-vs.-host disease (TA-GVHD) - prevention
irradiate components for:
-premature infants
-intrauterine or exchange transfusion
-stem cell or bone marrow transplants
-recipients of blood from a 1st-degree relative
-immunocompromised
-patients with leukemia or lymphoma
Delayed transfusion reactions - Nonimmunologic - iron overload - clinical signs
-diabetes
-cirrhosis
-cardiomyopathy
Delayed transfusion reactions - Nonimmunologic - iron overload - cause
build-up of iron in body
Delayed transfusion reactions - Nonimmunologic - iron overload - laboratory findings
increased serum ferritin
Delayed transfusion reactions - Nonimmunologic - iron overload - problem for?
patients receiving repeated transfusions over long period of time, e.g., patients with thalassemia, sickle cell anemia, other chronic anemias
Transfusion reaction investigation - signs & symptoms of possible transfusion reaction
-fever
-chills
-respiratory distress
-hyper- or hypotension
-back, flank, chest, or abdominal pain
-pain at site of infusion
-hives (urticaria)
-jaundice
-hemoglobinuria
-nausea/vomiting
-abnormal bleeding
-oliguria/anuria
Transfusion reaction investigation - specimens needed
-pre-transfusion blood
-post-transfusion blood
-post-transfusion urine
-segment from unit
-blood bag with administration set & attached IV solutions
Transfusion reaction investigation - immediate steps
-stop transfusion
-check all IDs & labels
-repeat ABO on post-transfusion sample
-visual check of pre- & post-transfusion samples for hemolysis
-DAT on post-transfusion sample; if positive, perform on pre-transfusion sample
Transfusion reaction investigation - signs of hemolytic reaction
-hemolysis in post-transfusion sample, but NOT in pre-transfusion sample
-mixed field agglutination in DAT on post-transfusion sample, but NOT on pre-transfusion sample
Transfusion reaction investigation - further steps if signs of possible hemolytic reaction
-check HGB in first voided urine after transfusion
-repeat ABO & Rh on pre- & post-transfusion samples & unit
-repeat antibody screen on pre- & post-transfusion samples
-AHG crossmatch with pre- & post-transfusion samples
Transfusion reaction investigation - additional tests that may be performed
-haptoglobin (decreased with hemolysis)
-gram stain & culture of unit
-bilirubin 5-7 hours after transfusion (sign of extravascular hemolysis)
-BUN & creatinine (sign of renal involvement)
Transfusion reaction investigation - reporting of transfusion-related fatalities
must be reported to FDA Center for Biologics Evaluation & Research (CBER) by phone or email ASAP, followed by submission of a written report within 7 days
Testing of Neonates (< 4 months) - ABO & Rh - specimen
cord blood, capillary, or venous blood
Testing of Neonates (< 4 months) - ABO & Rh - is ABO reverse grouping required?
no, ABO forward grouping only
Testing of Neonates (< 4 months) - ABO & Rh - how many times per admission?
once
Testing of Neonates (< 4 months) - antibody screen - specimen
serum or plasma of mother OR baby
Testing of Neonates (< 4 months) - antibody screen - required how many times per admission?
once
Testing of Neonates (< 4 months) - Crossmatch - specimen
serum or plasma from mother OR baby
Testing of Neonates (< 4 months) - Crossmatch - positive antibody screen
perform AHG crossmatch on units negative for corresponding antigen
Testing of Neonates (< 4 months) - Crossmatch - negative antibody screen
crossmatch not required
Hemolytic Disease of the Fetus and Newborn (HDFN) - Mothers at risk:
-ABO:
-Rh:
ABO: usually group O
Rh: Rh negative
Hemolytic Disease of the Fetus and Newborn (HDFN) - First child affected?
-ABO:
-Rh:
ABO: yes
Rh: not usually
Hemolytic Disease of the Fetus and Newborn (HDFN) - frequency:
-ABO:
-Rh:
ABO: common
Rh: uncommon
Hemolytic Disease of the Fetus and Newborn (HDFN) - Severity:
-ABO:
-Rh:
ABO: mild
Rh: can be severe
Hemolytic Disease of the Fetus and Newborn (HDFN) - DAT:
-ABO:
-Rh:
ABO: weak pos or neg
Rh: strong pos
Hemolytic Disease of the Fetus and Newborn (HDFN) - Spherocytes?
-ABO:
-Rh:
ABO: yes
Rh: rare
Hemolytic Disease of the Fetus and Newborn (HDFN) - Predictable?
-ABO:
-Rh:
ABO: no
Rh: yes (maternal antibody screen)
Hemolytic Disease of the Fetus and Newborn (HDFN) - Preventable?
-ABO:
-Rh:
ABO: no
Rh: yes (RhIG)
Rh Immune Globulin (RhIG) Workup - prenatal evaluation
-ABO & Rh (weak D not required) - if Rh positive, woman isn’t candidate for RhIG
-Antibody screen (confirm patient has not received prenatal RhIG, which may be detected in antibody screen) - if positive, ID antibody; if anti-D present, woman isn’t candidate
Rh Immune Globulin (RhIG) Workup - postpartum evaluation
ABO & Rh, including weak D, on baby:
–if baby is Rh neg = mother isn’t candidate
–if baby is Rh pos, draw mother’s blood after delivery & perform rosette test to screen for fetal blood
—Mother’s RBCs incubated with anti-D
—Anti-D coats fetal D-pos RBCs
—indicator D-pos RBCs added
—attach to anti-D on fetal D-pos RBCs, forming rosettes
–If rosette test pos, quantitate fetal bleed by flow cytometry or Kleihauer-Betke acid-elution test; fetal cells resist acid elution and stain pink; adult cells lose HGB and appear as “ghosts”
Rh Immune Globulin (RhIG) - composition
anti-D derived from pools of human plasma
Rh Immune Globulin (RhIG) - purpose
prevent immunization to D
Rh Immune Globulin (RhIG) - administration
-Antepartum: to Rh neg woman at 28 weeks of gestation
-Postpartum: within 72 hours of delivery when Rh-neg woman delivers Rh-pos baby
-Other obstetric events: to Rh-neg woman after spontaneous or therapeutic abortion, ectopic pregnancy, amniocentesis, chorionic villus sampling, antepartum hemorrhage, or fetal death
Note: May also be administered to Rh-neg recipients of Rh-pos blood or components
Rh Immune Globulin (RhIG) - Dose
1 dose per 15 mL of D-pos fetal RBCs (30 mL of fetal whole blood)
Calculating dose:
–if # to right of decimal point is >=0.5, round up to next whole # & add 1 vial, e.g., 1.6 vials calculated = 2 + 1 = 3
–if # to right of decimal point is <0.5, don’t round up; just use whole # & add 1 vial, e.g., 1.4 vials calculated = 1 + 1 = 2
Equipment/Reagent Quality Control - blood storage refrigerators & freezers, platelet incubators
system for continuous temp monitoring & audible alarm
Equipment/Reagent Quality Control - blood storage refrigerators & freezers, platelet incubators
system for continuous temp monitoring & audible alarm
Equipment/Reagent Quality Control - temperature recorder
compare against thermometer daily; calibrate as necessary
Equipment/Reagent Quality Control - alarms
check high & low temp of activation quarterly
Equipment/Reagent Quality Control - water baths
check temp daily
Equipment/Reagent Quality Control - heat blocks
check temp daily; periodically check each well
Equipment/Reagent Quality Control - centrifuges
-determine optimum speed & time for different procedures upon receipt, after repairs, & periodically
-check timer every 3 months, RPM every 6 months (with tachometer)
Equipment/Reagent Quality Control - cell washes
check tube fill level daily, AHG volume monthly; verify time & speed quarterly
Equipment/Reagent Quality Control - pipettes
calibrate quarterly
Equipment/Reagent Quality Control - antisera
test with pos & neg controls each day of use; use heterozygous cells for pos controls
Equipment/Reagent Quality Control - reagent cells
-check for hemolysis
-test each day of use with pos & neg controls
Equipment/Reagent Quality Control - reagent cells
-check for hemolysis
-test each day of use with pos & neg controls
Equipment/Reagent Quality Control - AHG
check anti-IgG activity each day of use by testing Rh-pos cells sensitized with anti-D
Which of the following enhancements allows antibodies and antigens to come closer together faster?
A. Albumin
B. LISS
C. Papain
D. PEG
B. LISS
Which of the following is utilized in weak D testing?
A. 22C incubation
B. treatment with LISS
C. 37C incubation and IAT
D. 37*C incubation only
C. 37*C incubation and IAT
Which of the following antibodies is/are often found as cold agglutinin(s)?
A. Anti-D and anti-C
B. Anti-K
C. Anti-M and anti-N
D. Anti-Fya and anti-Fyb
C. Anti-M and anti-N
Which of the following tests is required on donated blood per AABB and/or FDA?
A. Syphilis testing
B. Hepatitis testing
C. HIV testing
D. All of the above
D. All of the above
Select the preservative that will increase the shelf life of stored RBCs from 21 to 35 days.
A. Acid citrate dextrose
B. Citrate-phosphate-dextrose with adenine
C. Citrate-phosphate-dextrose-dextrose
D. Albumin
B. Citrate-phosphate-dextrose with adenine
What ABO type is capable of donating to any other ABO type?
A. AB negative only
B. A negative and B negative
C. O negative only
D. O negative and AB negative
C. O negative only
When a patient has a documented history of anaphylactic reactions, what should be done with the blood products before transfusion?
A. Wash the blood product
B. Irradiate the blood product
C. Only transfuse IgE-deficient blood
D. Wrm the blood product
A. Wash the blood product
Which of the following are typical laboratory findings in a patient suffering from a delayed hemolytic (extravascular) transfusion reaction?
A. Elevated serum bilirubin
B. Decreased haptoglobin value
C. Decreased HGB & HCT value
D. All of the above
D. All of the above
When performing an antibody ID, which of the following scenarios is expected if rouleaux is present?
A. Reaction with all cells in AHG, autocontrol positive
B. Reaction with all cells in AHG, autocontrol negative
C. Reaction with all cells at 37*C, neg in AHG, autocontrol positive
D. No reaction
C. Reaction with all cells at 37*C, neg in AHG, autocontrol positive
In which situation/condition is a 12-month donor deferral required?
A. Pregnancy
B. Travel to area endemic for malaria
-C. Previous whole blood donation
D. Recipient of influenza vaccine
B. Travel to area endemic for malaria