ASCP MLT BOC Immunohematology
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| What stimulates the Primary Immune Response? | First exposure to an antigen
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| How long is the lag phase for Primary Immune Response? | Days to Months
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| What type of antibodies are associated with Primary Immune Response? | IgM at first. May switch to IgG after 2-3 weeks (isotype switching)
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| What is the titer reaction for Primary Immune Response? | Rises slowly. Peaks then declines.
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| What does "lag phase" mean? | The period of inactivity after exposure to an antigen before the body produces an immune response.
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| What stimulates the Secondary Immune Response? | Subsequent exposure to antigen
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| How long is the lag phase for Secondary Immune Response? | Hours
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| What type of antibodies are associated with Secondary Immune Response? | IgG only
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| What is the titer reaction for Secondary Immune Response? | Rises faster and higher and stays elevated longer.
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| What is the structure and number of binding sites of IgG? | Monomer and 2 binding sites
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| What is the optimum temperature of reactivity for IgG? | 37°C
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| Which antibody (IgG or IgM) reacts in saline? | IgM
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| Which antibody (IgG or IgM) reacts best by the indirect antiglobulin test (IAT)? | IgG
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| What is the strength of complement fixation for IgG? | Moderate
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| Which antibody (IgG or IgM) causes transfusion reactions? | IgG
Not usually IgM, except ABO
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| Which antibody (IgG or IgM) crosses the placenta? | IgG
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| Which antibody (IgG or IgM) is destroyed by sulfhydral compounds? (dithiothreitol [DTT], 2-mercaptoethanol [2-ME] | IgM
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| What is the structure and number of binding sites for IgM? | Pentamer and 10 binding sites
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| Which antibody (IgM or IgG) is naturally occurring and which is Immune? | IgM is naturally occurring
IgG is Immune
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| What is the optimum temperature of reactivity for IgM? | 25°C or lower
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| What is the strength of complement fixation for IgM | Strong
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| What are the 2 signs of Antigen-Antibody reactions in blood bank testing? | Hemolysis and agglutination
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| What is the optimum pH for Ag/Ab reactions? | pH 5.5-8.5
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| What is LISS? | Low Ionic Strength Solution: lowers ionic stregth of suspending medium, allowing AG and AB to move closer together. Reduces incubation time to 10 minutes
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| What is prozone? | Too much antibody, not enough antigen, can cause false-negative
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| What is 20-30% albumin used for? | Causes agglutination by adjusting zeta potential between RBCs
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| What is zeta potential? | The difference in charge between the negative charged RBC surface and the cloud of positive ions that surround the RBCs. Reducing zeta potential allows RBCs to move closer together.
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| What is PEG? | Polythyleneglycol: Increases antibody uptake. Used for the detection and ID of weak IgG antibodies
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| What are enzymes used for in Blood Bank testing? | Reduces RBC surface charge by cleaving sialic acid molecules. M, N, S, Fya, and Fyb antigens are destroyed.
i.e. ficin, blomelin, and papain.
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| What is AHG? | Antihuman Globulin: Used to agglutinate cells that are sensitized (have antibody or complement attached)
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| Polyspecific (broad spectrum) AHG | Detects IgG and C3d
Used for DAT and in some labs, routine compatibility tests and anitbody detection
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| Monospecific AHG | anti-IgG only: detects clinically significant antibodies
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| What is the DAT? | Direct Antiglobilin testing: Detects in-vivo sensitization of RBCs by IgG antibody
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| What is the preferred specimen for DAT? | EDTA red cells
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| Which antiglobulin test (DAT or IAT) is incubation required? | IAT: Patient serum or plasma with reagent RBCs
or Patient RBCs with reagent antiserum
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| When is the DAT done? What is it's purpose? | Hemolytic disease of the newborn
Transfusion reactions
Autoimmune hemolytic anemia
Drug induced hemolytic anemia
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| What is the IAT? | Indirect Antiglobulin Testing: Detects in-vitro sensitization of RBCs by IgG antibody
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| What is the preferred specimen for IAT? | Serum, plasma, RBCs
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| When is the IAT done? What is it's purpose? | Antibody screen,
Crossmatch,
RBC phenotyping,
weak D testing
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| What is the A antigen's precursor and terminal sugar? | H substance and N-acetylgalactosamine
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| What is the B antigen's precursor and terminal sugar? | H substance and D-galactose
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| What is the frequency of ABO blood types in whites? | 45% O, 40% A, 10% B, 5% AB
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| What is the frequency of ABO blood types in Blacks? | 49% O, 27% A, 20% B, 4% AB
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| What is the frequency of ABO blood types in Hispanics? | 56% O, 28% A, 12% B, 4% AB
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| What is the frequency of ABO blood types in Asians? | 43% O, 27% A, 25% B, 5% AB
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| What is the genotype of Rh positive bood? | DD or Dd
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| What is the genotype of Rh negative blood? | dd
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| What is the frequency of D antigen in Whites and Blacks? | Whites 85%
Blacks 93%
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| R0 | Dce
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| R1 | DCe
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| R2 | DcE
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| Rz | DCE
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| r | dce
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| r' | dCe
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| r" | dcE
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| ry | dCE
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| Most common Rh genes in Whites (in decending order) | R1, r, R2, R0,r', r"
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| Most common Rh genes in Blacks (in decending order) | R0, r, R1, R2, r'
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| Which blood antigens are uncommon in Whites? (<25%) | K, Lea, Lua
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| Which blood antigens are uncommon in Blacks? (<50%) | K, Fya, Fyb, S, Lea, Lua, Lub
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| Which blood antigens are most common in Whites? (>80%) | k, Fyb, s, U, I, Lub
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| Which blood antigens are most common in Blacks? (>80%) | k, Jka, s, U, I, P1
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| I antigen presence | Much in Adult cells
Trace in cord
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| i antigen presence | Trace in Adult cells
Much in Cord cells
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| Naturally occuring antibodies | ABO, Lewis, P1, MN, Lua
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| Clinically significant Antibodies | ABO, Rh Kell, Duffy, Kidd, SsU
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| Warm antibodies | Rh, Kell, Duffy, Kidd
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| Cold antibodies | M, N, P1
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| Antibodies that usually only react with AHG | Kell, Duffy, Kidd
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| Which antibody can react in any phase of testing? | Lewis
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| Detection of antibody enhanced by enzyme treatment of test cells | Rh, Lewis, Kidd, P1
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| Antibodies not detected with enzyme treatment of test cells | M, N, S, Duffy
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| Which antibody is enhanced by acidification | M
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| Which antibodies show dosage? | Rh(not D), MNS, Duffy, Kidd
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| Which antibodies bind complement? | I, Kidd, Lewis
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| Which antibodies cause in vitro hemolysis? | ABO, Lewis, Kidd, Vell, and some P1
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| Which antibodies are labile in vivo and in vitro? | Kidd
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| What does labile mean? | deteriorates rapidly in storage
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| antibody that commonly causes anamnestic response | Kidd
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| Define anamnestic response | delayed transfusion reaction
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| Which antibody is associated with paroxysmal nocturnal hemoglobinuria? | Anti-P
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| What is paroxysmal nocturnal hemoglobinuria PNH? | Defect in RBC membrane, makes in more susceptible to hemolysins in an acid environment. Hgb found in urine after periods of sleep.
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| Which antibody is associated with cold agglutinin disease and Mycoplasma pneumoniae infections | Anti-I
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| Which antibody is associated with infectious mononucleosis? | Anti-i
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| Antibody ID Reaction: same strength and in one phase only | Suggestive of single antibody
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| Antibody ID Reaction: Varying strength | Multiple antibodies, antibody exhibiting dosage, antigens of differing strength
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| Antibody ID Reaction: in different phases | Combination of warm and cold antibodies, antibody with wide thermal range
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| Antibody ID Reaction: All cells in AHG, autocontrol negative | Multiple antibodies, antibody with high frequency antigen
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| Antibody ID Reaction: All cells in AHG, autocontrol positive | Warm autoantibody
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| Antibody ID Reaction: All cells at 37°C, negative in AHG, autocontrol positive | Rouleaux
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| How long do you retain patient specimen and unit segment and what temperature do you store it at? | 7 days following transfusion at 1-6°C
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| How many days before a transfusion must the specimen be collected for compatability testing? | 3 days
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| What is an antiglobulin crossmatch? | Recipient serum and donor RBCs are tested with the IAT. Required when recipient has, or previously had, a clinically significant antibody
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| What is an abbreviated crossmatch? | Recipient serum and donor RBCs are tested in immediate spin only.
Permissible if recipient doesn't have, and has never had, clinically significant antibodies. Test of ABO compatibility.
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| What is a computer crossmatch? | Computer check of donor ABO and Rh type and recipient ABO and Rh type. Permissible if recipient doesn't have, and has never had, clinically significant antibodies.
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| What tests are done on newborns for transfusion? | ABO and Rh typing (forward grouping only), Antibody screen,and crossmatch (only need if AB screen is positive)
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| What conditions must be met if RBCs are being used for reissue? | 1-10°C, closure not broken, at least one segment left, unit inspected, records indicate blood has been reissued
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| Clinical signs of hemolytic (intravasuclar) txrxn | Fever, chills, shock, renal failure, DIC, Pain in chest, back, or flank
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| Clinical signs of hemolytic (extravascular) txrxn | Fever, anemia, mild jaundice 2 or more days after tranfusion
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| Clinical signs of febrile txrxn | Increase of temperature > or = to 1°C within 24 hours of transfusion with no other explanation
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| Clincal signs of allergic txrxn | Hives
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| Clinical signs of anaphylactic txrxn | Bronchospasms
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| Clinical signs of circulatory overload txrxn | Coughing, cyanosis, difficulty breathing
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| Clinical signs of septicemia txrxn | Fever, cramps, diarrhea, vomiting, muscle pain, DIC, shock, renal failure
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| Specimens needed for a txrxn investigation | Pre-transfusion blood
Post-transfusion blood
Post-transfusion urine
Segment from unit
Blood bag with administration set and attached IV solutions
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| At what gestational age is RhIG administered? | 28 weeks
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| How long postpartum is RhIG given to Rh negative women with Rh positive baby? | within 72 hours
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| What is the dose of RhIG given to Rh negative mother with Rh positive baby? | One dose per 15 ml of D postive fetal RBCs.
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| AABB age requirement for blood donation | At least 17 yrs old
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| AABB weight requirement for blood donation | At least 110 lbs to donate 525 ml
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| How often can you donate blood (AABB)? | Every 8 weeks
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| AABB blood pressure requirement | Systolic < or =180
Diastolic < or =100
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| AABB pulse requirement | 50-100 with no pathological cardiac irregularities
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| AABB hemoglobin/hematocrit requirement | > or = 12.5 g/dL HGB
> or = 38% HCT
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| AABB body temp requirement | < or = 37.5°C (99.5°F)
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| 3 day blood donor deferral (AABB) | Apirin containing medicaitons if donor is sole source of plts
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| 2 week donor deferral (AABB) | Measles, mumps, polio, or yellow fever vaccines
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| 4 week donor deferral (AABB) | Rubella vaccine
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| 6 week donor deferral(AABB) | Pregnancy
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| 12 months donor deferral (AABB) | Syphilis, Gonorrhea, Animal bite, HBIG, Tattoo, Mucous membrane exposure to blood, Needle stick, Household or sexual contact with individual with hepatitis, sexual contact with individual with HIV, Traval to area endemic with Malaria
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| 3 year donor deferral (AABB) | Malaria, or from an area endemic for malaria
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| Permanent donor deferral (AABB) | Parenteral drug use, family history of Creutzfeldt-Jakob disease, Treated with growth hormone, Viral hepatitis after 11th bday, Postive HBsAg, Repeatedly reactive anti-HBc, anti-HCV, anti-HTLV, or anti-HIV,Babesiosis, Chagas' disease
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| Volume of anticoagulant for blood unit collection | 63 ml
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| Volume of blood collected for blood unit | 450 +-45 ml
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| Low volume unit | 300-404 ml; label "low volume unit" don't use to prepare platelets or plama components
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| When should the volume of anticoagulant be reduced? | When the blood volume is <300 ml
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| Time of collection for blood unit | 4-10 minutes, if >15 minutes, blood may not be suitable for preparation of plts or FFP
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| Temperature of blood unit | 20-24°C for up to 8 hours if components are going to be prepared, otherwise 1-6°C
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| Acid-citrate-dextrose | ACD anticoagulant
shelf life 21 days
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| Citrate-phosphate-dextrose | CPD Anticoagulant
shelf life 21 days
High pH preserves 2,3-DPG better. Better O2 delivery.
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| Citrate-phosphate-dextrose with adenine | CPDA-1
shelf life 35 days
Adenine increases adenosine disphosphate ADP
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| Adsol | AS-1 Additive
Shelf life 42 days
Provides nutrients for improved viability
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| Nutricel AS-2 | AS-2 Additive
Shelf life 35 days
Provoides nutrients for improved viability
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| Nutricel AS-3 | AS-3 additive
shelf life 42 days
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| Phosphate-inosine-pyruvate-adenine | PIPA rejuvinating solution
used to salvage rare or type O units up to 3 days beyond expiration, must then be transused within 24 hours or frozen
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| Donor serological testing (AABB or FDA) | ABO, Rh, AB Screen, RPR, HBsAg, Anti-HBc, Anti-HCV, Anti-Hiv 1/2, HIV-1-Ag, Anti-HTLV-1 and 2
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| Autologous donation frequency | every 3 days but not within 72 hours of transfusion
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| Autologous donation Hgb requirement | not less than 11 g/dL
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| Temperature and shelf life of RBCs | 1-6°C and 35 days in CPDA-1, 42 days in AS-1
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| Temperature RBCs frozen | 40% glycerol: < or = -65°C
20% glycerol: < or = -120°C
1-6°C after glycerolization
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| Shelf life or RBCs frozen | 10 years after phlebotomy,
24 hours after deglycerization
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| Temperature and shelf life of washed RBCs | 1-6°C
24 hours after washing
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| Temperature and shelf life of Leukocyte reduced RBCs | 1-6°C
Closed system:10 years
Open system: 24 hours
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| Temperature and shelf life of Rejuvinated RBCs | 1-6°C
24 hours after rejuvination if not frozen
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| Temperature and shelf life of irradiated RBCs | 1-6°C
Original outdate or 28 days from irradiation, whichever occurs first
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| Temperature and shelf life of FFP | < or = -18°, After thawing 1-6°C
12 months, After thawing transfuse within 24 hours
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| Temperature and shelf life of Cryoprecipitate | < or = -18°C, After thawing room temperature
12 months, After thawing transfuse within 6 hours if unit is not entered, within 4 hours in pooled
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| Temperature and shelf life of platelets | 20-24°C
5 days
5 days with agitation
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| Temperature and shelf life of granulocytes | 20-24°C
24 hours
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| Increased in stored blood | Plasma K+, Plasma NH3, Plasma Hgb, microaggregates
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| Decreased in stored blood | pH, ATP, 2,3-DPG, Viable RBCs, WBCs, and platelets, Labile coagulation factors
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| Centrifuge QC | Check RPM and timers quarterly
Determine optimum speed and time for procedures upon receipt, after repairs, and semiannually.
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| Cell washers QC | Check volume of saline and AHG in each tube,
verify time and speed of centrifugation periodically
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| Waterbaths QC | Check temperature daily
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| Heat blocks QC | Check temperature daily. Periodically check each well.
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| Refrigerators QC | System to monitor temperature continuously and to record temperature at least every 4 hours. Alarm system with audible signal. Must be 1-6°C
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| Freezers QC | System to monitor temperature continuously and to record temperature at least every 4 hours. Alarm system with audible signal.
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| Alarms QC | Check high and low temperatures of activation quarterly
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| Platelet incubators QC | System to monitor temperature continuously and to record temperature at least every 4 hours. Should be 20-24°C. Check RPM periodically.
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| Pipettes and droppers QC | Determine average delivery volume. Calculate # of drops that will give 80:1 serum to cell ratio
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| Antisera QC | Test with positive and negative controls each day of use. Use heterozygous cells for positive controls
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| Reagent cells QC | Check for hemolysis. Test daily with pos and neg controls
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| AHG QC | Check anti-IgG activity by testing Rh-positive cells sensitized with anti-D
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| How long should QC records be kept? | 5 years or longer
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| When are washed RBCs needed? | Anemic patients with paroxysmal nocturnal hemolgobinuria, ABs to IgA, or history of febrile reactions
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| When are leukocyte reduced RBCs needed? | Anemia with history of febrile reactions
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| When are irradiated RBCs needed? | Intrauterine transfusions, immunocompromised recipients, recipients of blood from a relative, or bone marrow transplant patients
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| When is FFP needed? | Deficiency of coagulation factors
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| When is cryoprecipitate needed? | Hemophilia A, Von Willebrand's disease, hypofibrinogenemia, Factor XIII deficiency
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| When are platelets needed? | Prevent or stop bleeding in patient with thrombocytopenia or abnormal platelet function.
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| When are granulocytes needed? | Neutropenia with infection
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| How long after receiving a donation of blood products are patients deferred for donation? | 12 months
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| CPDA-1 ingredients | Citrate, Dextrose, Adenine, Sodium biphosphate
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| Citrate purpose in CPDA-1 | Prevents coagulation by chelating calcium
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| Dextrose purpose in CPDA-1 | Supports adenosine triphosphate (ATP) generation
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| Adenine purpose in CPDA-1 | Substrate for ATP synthesis
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| Sodium biphosphate in CPDA-1 | Prevents excessive fall in pH during storage
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| Blood product that must be separated from whole blood and frozen within 8 hours of collection | Fresh Frozen Plasma
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| How is cryoprecipitate prepared? | By thawing FFP between 1-6°C, removing plasma, and freezing within 1 hour.
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| RBCs must be frozen in: | glycerol within 6 days of collection
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| How long after plasmapheresis must a person wait to donate a unit of Whole Blood? | 48 hours
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| Each unit of whold blood will yield approximately how many units of cryoprecipitated AHF? | 80 units
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| 90% of all platelet pheresis units tested contain how many platelets per ul? | 3.0 x 10"
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| Terminal sugar for H antigen | L-fucose
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| Approximate % of original plasma content of Factor VIII recovered in Cryoprecipitated AHF | 40-80%
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| What is the formula to calculate fetomaternal hemorrhage for RhIg? | (fetal cells/number of cells counted)x maternal blood volume
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| Liley method of predicting the severity of HDN is based on aminiotic fluid: | change in optical density measured at 450 nm
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