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immunohematology Flashcards

MLT certification exam

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984499619Collection of donor blood- Follows registration, medical history, and physical examination - Preparation of venipuncture site will minimize the risk of bacterial contamination: drawing will be with aseptic techniques - Scrub site for minimum 30 seconds0
984499620Maximum collectionNo more than 10.5 mL of whole blood per kilogram of body1
984499621Medications evaluated by MDPlatelets from donors who have taken medication within 48hrs that irreversibly affect platelet function (aspirin) must not be used as the Only source of platelets (can be part of a pool of platelets)2
984520482criteria for donor selection-Age: >16 yrs minimum or conform to state law, no max -Temp: ≤37.5°C or ≤99.5 °F -Blood pressure: ≤180 systolic and ≤100 diastolic -Hgb/Hct: ≥ 12.5g / ≥ 38% -Pulse: 50-100 beats/min -Weight: minimum 110lb/ 50 kg3
984520483ACD/CPD/CPD2Anticoagulant 21 day expiration4
984520484CPDA-1Anticoagulant 35 day expiration5
984520485Additive solutions42 day expiration6
984520486Rejuvenating solution- Restores 2,3-DPG and ATP - Can freeze unit or if used in 24hrs, can be stored at 1-6C - must wash cells before transfusion to remove solution7
984520487Autologous DonationsDonations to self: no age limit Hct - 33%, Hbg- 11 g/dL No bacteremia Preoperative collection must be labeled for autologous use only and used only for this patient Autologous units - segregate from allogeniec units8
984558791Low volume collectionAutologous donations -use regular blood bags; volume drawn <10.5 mL/kg body weight for minimum weight (450 + 45 mL plus testing sample) - If 300-404 mL drawn, label as Red Blood Cells Low Volume (components may not be made from these units) - If < 300 mL drawn, use proportionately less anticoagulant9
984558792Example low volume donor70lb/110l X 450ml (standard donation) = 286 ml -If donor weight in kg, divide weight by 50 instead of 110 and then multiple by 450 -Amount of anticoagulant to use: 286 X 14% = amt of anticoag 286 X .14 = 40; 40ml -Amount of anticoagulant to take out 63-40 = 23ml: remove 23ml of anticoagulant from primary bag into attached satellite bag prior to draw10
984558793Blood salvageinoperative collections "salvaged" blood collected during surgery, washed on-site and returned to patient during procedure11
984671866Apheresis termsCytapheresis - Cells Plasmapheresis- plasma Plateletpheresis - platelets Leuka/granulocytapheresis - Leukocytes/granulocytes12
984671867Hemapheresis/Apheresis collection-Donor criteria same as for whole blood - limited number of donor exposure - Apheresis instruments can selectively remove needed component and return components not needed.13
984671868CytapheresisPlatelets, granulocytes and leukocytes - donations at least 2 days apart and no more than 2 days in any 7 days - If RBC cannot be returned, must wait 8 weeks RBCs - A two unit RBC donation must not decrease donors hematocrit below 30% or hemoglobin below 10g/dl Hematopoetic progenitor cells - Can be collected from peripheral circulation14
984671869Therapeutic-Removal of pathogenic substance (platelets, leukocytes, plasma substances) - Immunoadsorption removes specific substances from plasma via antigen-antibody or chemical reactions (IgG or immune complexes)15
984671870Hematopoeitic progenitor and stem cells- used to reconstitute bone marrow post chemotherapy/irradiation or to replace abnormal marrow cells with normal marrow cells (congenital immune deficiencies, anemia's, malignant disorders of bone marrow, red cell disorders) - Cells obtained from bone marrow, umbilical cord blood and peripheral blood (apheresis) -Allogeneic marrow - HLA - identical match lowers GVHD risk; ABO compatibility not required16
984671871Tests performed on donor blood*ABO - Resolve discrepancies *Rh - Weak D determination on D negatives *Antibody screen - Clinically significant antibodies: FFP cannot be prepared from these units: platelets and cryoprecipitate can (contain minimum plasma) *Syphilis- Serologic test such as RPR *Vial diseases - HBsAG, Anti-HBc, Anti-HCV, Anti-HIV 1/2, Anti-HTLV-I/II and NAT for HIV-1 RNA, HCV RNA and West nile RNA17
984671872Whole blood-Given in cases of severe shock (blood loss ≥ 25% blood volume) needing rbc's for oxygen and plasma for volume -Rarely used due to increased use and availability of components18
984671873Red Blood cells (packed cells)- Red cells with plasma removed - Provides same oxygen carrying capacity as whole blood with less volume - < 80% hct (indicates sufficient plasma removal); 55-65% hct if additive solution used - 1 unit raises hemoglobin - hbg - 1g or hematocrit -hct- 3%19
984671874Washed red cells- Plasma removed by successive saline washes (automated instruments) - Prevents allergic response to plasma proteins and anaphylactic shock in IgA deficient patients with anti-IgA (IgA is in normal plasma) - Expires 24 hours after seal of original unit broken20
984671875Leukocyte reduced red cells- 85% of red cells retained - Final wbc count < 5 X 10^6 to prevent febrile nonhemolytic reactions and for other uses ( to prevent CMV transmission) - Preparation by filtration preferred: washing will remove leukocytes also - Used primarily for patients with repeated febrile nonhemolytic (FNH) reactions; Usually due to presence of cytokines released from white cells or alloimmunization to HLA or leukocyte anitgen21
984687666Frozen cells/ Deglycerolyzed cells- Cells protected from ultra low temperatures by cryoprotective agent (40% glycerol) - Must be thawed at 37°C and glycerol removed prior to transfusion - 80% of original red cells must be recovered - Used for storage of autologous units and "rare" units - expire in 10 years - Stored at ≤ -65; 1-6°C for 24Hours after deglycerolizing (open system)22
984687667Fresh Frozen Plasma (FFP)-Prepared by separating cells and plasma by centrifugation and freezing plasma within 8 hours of collection -Expires 1 year from date of collection when stored at ≤ -18°C or 7 years stored at ≤ -65°C -Once thawed (between 30-37°C), expires in 24 hours, if stored at 1-6°C - Used for multiple coagulation deficiencies, factor XI deficiency and other congenital deficiencies for which no concentrate is available23
984687668Cryoprecipitate (cryoprecipitated antihemophilic factor)- When FFP frozen within 8 hours of whole blood collection is thawed at 1-6°C, a cold insoluble portion of plasma forms - CRYO - CRYO- is separated from thawed FFP and refrozen within one hour - Must contain ≥150mg of fibrinogen and ≥80 IU/bag of factor VIII - Also contains vWF, facor XIII and fibronectin - Store at ≤ -18°C for 1 year from date of phlebotomy: RT after thawing -tranfuse within 6 hours of thawing; 4 hours after pooling24
984695876Cryoprecipitate used for?-Fibrinogen and Factor XIII deficiencies - Severe von Willebrand disease ( some factor VIII concentrates contain vWF) - Topical fibrin sealant - Seldom used for hemophilia because of factor VIII concentrates which have little or no risk of viral infection transmission25
984695877Factor Concentrates/Recombinant products- Concentrates are prepared from plasma pools that are tested by NAT and processed to inactivate and remove pathogens - ↑ levels of specific factors with minimal volume compared to FFP - Hemophilia A - treat severe with recombinant Factor VIII and mild with DDAVP (stimulates endogenous Factor VIII release) - Hemophilia B - treat with recombinant Factor IX - Inhibitors to Factor VIII- recombinant Factor VII (rFVIIa) approved for use to bypass Factor VIII - von Willebrand disease - treat with factor VII concentrates that have vWF; Mile cases treat with DDAVP26
984695878Platelets preparation from whole blood-Prepared from whole blood (stored at 20-24°C prior to precessing) - First a light spin- to remove red cells, followed by heavy centrifugation - to spin down platelets and white cells -Supernatant plasma is expressed into another bag and may be frozen (FFP) - Remaining plasma, platelets and WBC = platelets27
984820381Platelet conditions-For severe thrombocytopenia and platelet dysfunction - Prophylactic use of platelets when platelet count is low is controversial - threshold depends on patient's risk of bleeding - Contraindicated in TTP and heparin-induced thrombocytophenia28
984820382Platelet donor on drugsPlatelets from donors who are within 48 hrs of taking drugs (aspirin) that impair platelet function should not be used as a single source.29
984820383Platelet refractorinessLack of expected response - Antibodies to HLA class I antigens - Platelet antibodies or neutrophil/lymphocyte antibodies30
984820384Platelet transfusion- 1 unit of platelets should raise platelet count 5,000-10,000 in average sized adult -Do not transfuse through a microaggregate filter - Only one ABO type/pool: expires 4 hours after pooling in an open system31
984820385Platelet QC-pH ≥ 6.2 at end of storage: stored in volume of plasma necessary to maintain pH, usually 30-70 cc - > 5.5X10^10 platelets/unit in 75% or > 3X10^11 platelets/plateletpheresis in 90% of units tested - Store continuously rotating at 20-24° C RT - Outdate depends on type of bag used - Individual leukoreduced platelets- <8.3X10^5 leukocytes; Leukoreduced pooled platelets or plateletpheresis product - <5X10^6 leukocytes - Must be checked for bacteria before issuing32
984820386Infant platelet transfusionTransfuse ABO compatible to infant33
984820387D pos plateletsMay have some residual RBC's - consider administering RhIg to D neg women of childbearing age who have received D pos platelets34
984820388Grandulocytes-Usually obtained by apheresis - Decline in use due to new antibiotics, recombinant growth factors and adverse effects from granulocyte transfusion (acute lung injury) - used for neutropenic patients with documented gram negative sepsis who have not responded to antibiotics - Can transmit CMV, induce HLA immunization and cause GVHD, if not irradiated - Stored without agitation at 20-24°C for up to 24hrs, but should be transfused ASAP - ABO-compatible with recipient35
984854476irradiated blood and components-Prevents graft (donor lymphocytes) vs. host disease (GVHD) by inactivating donor lymphocytes - Recommended for anyone at risk of transfusion associated GVHD; fetus receiving intrauterine transfusion; donor is blood relative of recipient; donor is HLA matched - Minimum of 25 Gy (2500 cGy) - RBC's expired on original outdate or 28 days after irradiation, whichever is first36
984854477How much anticoagulant/bag?63ml37
984854478What is expiration dates based on?Expiration based on expectation of 75% of transfused cells will be in circulation 24hrs after transfusion38
984854479What happens to plasma while in storage?↑ NH4 and K+ ↓ pH and Na+39
984854480Transporting blood and components- Red cells kept at 1-10°C - Platelets and granulocytes kept at 20-24°C - Frozen components kept frozen40
984854481Expiration of blood/components when seal is broken-Products stored at 1-6°C = 24hrs - products stored at 20-24°C = 4hrs41
984854482Pooling components-If red cells visible in pooled product, patient plasma antibodies should be compatible with those red cells - Expiration of pooled components a. Platelets- 4hrs - open system b. Cryoprecipitate- 4 hrs - open system42
984854483Return of unit bloodUnit blood cannot be returned and reissued if >10°C (RT 15-30min) or if seal disturbed43
984868158What are ABO antigensImmunodominant sugar on cell surface44
984921315Subgroups of AA1 and A2 - Principle subgroups of A - Serological difference based on reactivity with anit-A1 (dolichos biflorus or human anti-A1). Dolichos biflorus- lectin - plant or seed extract diluted to agglutinate specific human blood group antigens) - A1 cells are agglutinated - A2 cells are not agglutinated Other subgroups - A3, Ax - contain less A antigen and more H antigen45
984921316Bombay PhenotypeLack of H is genetically hh = bombay - hh has no fucose which is needed for attachment of A or B sugars - cell types as O - Anti-H lectin (Ulex europaeus) will not agglutinate bombay cells (hh) but will agglutinate O cells (Hh or Hh)46
984921317Se geneSecretor - allows expression of A, B, H and Leb in body fluids47
984921318Le antigensLe antigens are plasma antigens which adsorb onto red cells as individuals mature - Individual with H and Le (but not Se gene) genes will have H and Lea on red cells and Lea only in saliva- Se not needed for presnece of Lea in saliva but it is for H - Individual who has h, Se and Le genes will have H and Leb on the red cells and H, Leb and decreased amounts of Lea in the body fluids48
984921319Cell typing- Unknown cells + antisera = no agglutination a. Cells lack antigen to which antisera (reagent antibody) corresponds - Unknown cells + antisera = agglutination a. Cells possess antigen to which antisera corresponds49
989049291Serum TypingReverse typing - Testing is performed at RT with saline suspended known group A1 and B red cells - Optimum reactivity of serum anti-A and anti-B is 4C -Unknown serum + reagent red cells = No agglutination a. Serum lacks antibody to antigen on red cell - Unknown serum + reagent red cells = Agglutination a. Serum has antibody to antigen on red cells50
989049292Rouleuax (red cells)Failure to wash cells - Repeat with saline washed cells51
989049293Mixture of cell types (red cells)-Example - A or B transfused with O cells - Check transfusion history52
989049294Subgroups (red cells)- A2 with or without anti-A1 - Test with Anti-A1 for A subgroups53
989049295Unusual Genotypes (red cells)- Bombay - Test with Anti-H for bombay (not agglutinate) Bombay serum will agglutinate A1 and B cells as well as Group O screening cells54
989049296Disease processes (red cells)- Leukemia or bacteria (acquired B phenomenon) - Check patient diagnosis - Id beyond scope of this review55
989049297Rouleaux (serum)- Due to increased serum proteins - Waldenstrom's or multiple myeloma - Saline replacement56
989049298Room Temp or cold reacting antibody- H, I, M, N, P1 or lewis or anti-A1 in an A2 or A2B individual - React with corresponding antigens on reverse cells - Mini cold screen or panel (test at lower temps)57
989049299Age- Elderly (antibody production has decreased) - Newborn (antibody production has not reached optimum levels - Missing antibodies - check patient age, Mini cold panel (may enhance serum Anti-A or B so interpretation will agree with cell grouping)58
989049300Comprised immune system- Hypogammaglobulinemia - Check patient diagnosis - Mini cold panel59
989049301Saline replacementPrinciple: Differentiate rouleaux (aggregation) from agglutination. - Roulauex = stack of coins60
989049302Antigens of Rh systemD, C, E, c and e - D is the only antigen routinely tested61
989068398D antigen- most immunogenic of all blood group antigens - D typing (Rh typing) is based on presence or absence of D when tested with anti- D62
989068399Weak D-D reactive at antiglobulin phase only - Must have negative D control to be valid - Weak D positive is considered D positive - Tested for weak D required on donor and OB patients63
989068400Monoclonal/Polyclonal anti-D- Separate D control not needed for A, B or O positive cells - A negative reaction with reagent anti-A and or Anti-B in patient cell typing is the negative control - D control needed for any AB positive and for any immediate spin negative cell carried through to AHG for weak D typing - If patient is AB positive, must use a 6-8% albumin control, autocontrol or DAT (patient A and B cells typed with reagent anti-A and anti-B will be positive in an AB positive patient - Most common cause of positive D control is a positive DAT64
989068401High protein anti-D- Generaly replaced by monoclonal or monoclonal/polycolonal blend reagents - If used, Must use D control produced by same manufacturer as the anti-D65
989068402Unusual phenotypesRh null - no D, C, E, c or e antigens - Cells have associated hemolytic anemia since Rh structure is integral part of Rbc membrane66
989068403Rh antibodiesIgG clinically significant - May agglutinate at 37C as well as AHG - Anti-C, -c, -E, -e react stronger with enzyme-treated cells67
989068404IgM antibodiesAnti- I, -H Anti-M, -N Anti-P1 Anti-Lea, -Leb68
989068405IgG antibodiesAnti-D Anti-C, -c Anti-E, -e Anti- M (some) Anti-K, -k Anti-Fya, -Fyb Anti-Jka, -Jkb69
989076498Lewis antigens-Plasma antigens that adsorb onto RBC's - Lea and Leb are not alleles - Not on cord blood - Antibodies a. Do not cause HDN (lewis antigens are not on fetal cells and lewis antibodies usually IgM) b. IgM antibody - can be hemolytic c. Usually only seen in Le(a-b-) persons d. Often seen in pregnant women who may temporarily become Le(a-b-)70
989076499I, i antigensI - absent or weak on cord cells - i converts to I as infant matures due to branching of carbohydrate chians - I and i are not alleles - Antibodes a. IgM cold antibody b. Reacts with all adult cells (except rare i adult) c. May mask clinically significant alloantibody d. Remove anti-I to detect underlying antibodies by: -An autoadsoption (recent transfused) or allogeneic adsorption - RESt adsorption - Prewarming serum and using IgG AHG instead of polyspecific71
989076500P antigen- P1 antigen strength deteriorates upon storage - Antibodies a. Anti-P1 -IgG cold antibody - Anti-P1 can be neutralized to reveal other clinically significant alloantibodies (P1 substance in hydatid cyst fluid) b. Anti-P - Autoantibody-IgG; Donath-Landsteiner biphasic antibody found in paroxysymal cold hemoglobinuria - Reacts with all P or P1 positive cells72
989296628MNSs antigensM/N and S/s are both codominant alleles -Anit-M and -N a. Usually cold IgM, No HDN b. Often show dosage (property whereby antibody reacts strongest with cells having a homozygous expression of antigen as opposed to heterozygous cells -Anti-M a. Many examples are IgG and can cause HDN b. May require acidification of serum to identify -Anti-S and anti-s a. IgG -Anti-U a. IgG b. Formed by african american individuals who lack S, s and U73
989588095Kell- K and k are codominant alleles - 91% are K negative - Antigens inactivated with 2-ME, DTT or AET - IgG74
989588096Kidd- Jka and Jkb are codominant alleles -IgG - React stronger with enzyme treated cells - Titers rise and fall rapidly - Associated with delayed transfusion reactions - Often show dosage75
989588097Duffy-Fya and Fyb are codominant alleles - 68% african americans are Fy(a-b-) - Antigens destroyed by enzymes - Antigen typing- Fy(a+b-) -Caucasians - homozygous for Fya (FyaFya) - African americans - Probably heterozygous for Fya (FyaFy-)- may show dosage76
989588098Paternity testingMaternity is assumed There is a chain of sample custody that must be adhered to in legal cases Molecular techniques are replacing serological methods -RBC blood groups with codominant alleles can be used for parentage testing along with HLA system and DNA analysis77
989841761Direct paternity testing-Direct exclusion - marker present in child, absent from father and mother78
989841790Indirect paternity testingIndirect exclusion - Child lacks a marker that the alleged father must transmit79
989841804enhancement mediaAlbumin LISS Enzymes polyethylene glycol80
989841807Albumin enhancement media-Bovine - ↓ net negative surface charge - Only ↑ antibody uptake if under low ionic conditions - Rh antibodies may show at 37°C81
989841810Low Ionic Strength Saline (LISS) enhancement media- ↑ antibody uptake which allows ↓ in incubation time82
989841812Enzymes enhancement media- bromelin, ficin, papain and trypsin - removes sialic acid which ↓ negative surface charge and promotes cell agglutination - ↑ reactivity of Rh, Kidd and Lewis antibodies - Usually ↑ warm and cold autoantibodies - Destroys M, N, S, Fya and Fyb antigens83
989841813Polyethylene glycol (PEG) enhancement media- ↑ antibody uptake - removes water which ↑ antibody concentration which promotes antibody uptake84
989841826DAT1. Antiglobulin added to 3-4 times washed cells 2. If cells coated in vivo, antiglobulin will react with the IgG antibody and/or complement (depending on type of AHG used) 3. EDTA sample is optimum - EDTA chelates CA++ preventing complement activation by plasma antibody (causes false + DAT) 4. Add check cells to all negative antiglobulin tests in antibody detection and compatibility testing85
989919392AHG reagents- Can be polycolonal, monoclonal or blends monocolonal or monocolonal/polycolonal - Polycolonal = Inject animal with purified IgG, IgA, IgM, C3 or C4 - Monoclonal = Hybridoma derived86
989919393Polyspecific AHGAntibody to human IgG and C3d component of complement - Other complement components may be present87
989919394Monospecific AHGAntibody to IgG or to C3b, C3d88
989919395AHG reagents cont.-Perform DAT with polyspecific to screen and monospecific to characterize the globulin - Perform IAT with monospecific anti-IgG to avoid cold, complement-binding antibodies -Use check cells to confirm all negative antiglobulin test in antibody detection and compatibility testing when using anti-IgG a. Confirms AHG added and not neutralized (insufficient removal of serum proteins prior to addition of AHG)89
989919396Elution principle- based on breaking antigen-antibody bound, removing antibody from cell surface - Used to determine antibody specificity in cases of positive DAT due to IgG antibodies (HDN and transfusion reactions)90
989919397Types of elutions-Lui freeze - thaw and heat - ABO antibodies - Low pH acid, digitonin-acid, cold acid, and dichloromethane - all antibodies - no single method best for all antibodies91
989919398Last wash control - elutions1. Prior to elution, red cells coated with antibody should be thoroughly washed to remove any residual serum antibody 2. Test "last wash" (supernatant) before performing elution or in parallel with eluate 3. "last wash" should show no reactivity with reagent cells 4. Positive test results using "last wash" indicate serum antibody contamination of supernatant- if performed before elution, wash agian- if performed in parallel, test invalid - repeat92
989919399Neutralization (inhibition) tests1. Soluble antigen can bind with antibody to inhibit a reaction with RBCs - allows detection of alloantibodies "masked" by the following antibodies a. Lewis substances - in saliva b. P1 substance - in hydatid cyst fluid and pigeon egg whites c. Sda substance - most abundant in urine d. ABH sugars- inhibit anti-A, -B, -H e. Chido and rodgers substances - epitopes of C4 (complement)93
989919400InactivationSulfhydryl reagents - AET and DTT - destroys or weakens Kell system - ZZAP - enzyme + DTT -- destroys Kell system and those systems destroyed by enzymes - DTT and 2-ME - destroys or diminishes activity of IgM antibodies94
990017269Adsorption-Used to a. Separate multiple antibodies b. Remove autoantibody- reveal alloantibody "masked" by autoantibody c. Confirm antigen existence on RBC d. Confirm antibody specificity -Autologous adsorption (patients own serum and cells) can be used for patients not recentlry transfused - Allogeneic adsorptions (patients serum and other cells) can be used on patients recently transfused95
990017270Gel testing - Column agglutination- Unagglutinated cells pass through gel - agglutinated cannot - Negative = cells at the bottom - Positive = The cells remain on the top or partially travel through the gel depending on agglutinate size -Phenotype cells when reagent antisera is in cell- agglutinate when exposed to antibody during centrifugation - Antiserum/Serum/Plasma and cells can be added together in the upper chamber - sensitized cells agglutinate when exposed to the IgG in the gel and cannot go through - DAT by gel- No washing needed - Only RBCs go through gel and sensitized cells agglutinate when exposed to IgG in the gel96
990049363Solid Phase- Antibody or antigen - fixed to a microwell plate a. Antibody fixed to plate * RBCs with the antigen are added * Adhere to antibody on sides of microplate b. Antigen fixed to plate * Plasma with the antibody is added * Antibody will adhere to antigen on sides * Wash: add check cells to attach to antibody - Positive reactions have cells adhering to sides of microwell plate - Negative reactions have RBC pellet at bottom of plate since no attachment97
990049364Pretransfusion testing samples- Serum or plasma from intended recipient - Labeled with 2 unique identifiers and date of collection - must have system to identify phlebotomist - Retain for a minimum of 7 days after transfusion98
990049365Pretransfusion tests- ABO and D grouping - Antibody screen - Crossmatch - Autocontrol not required - Compare current results with prior testing99
990049366Crossmatch-Patient serum mixed with donor RBCs - Observe for agglutination or hemolysis - Demonstrate ABO compatibility - Carry through to 37C incubation with AHG if current antibody screen positive or prior history of clinically significant antibodies100
990049367Immediate spinImmediate spin or electronic (computer) crossmatch if current antibody screen negative and no prior antibody history101
990049368Are antibodies required to be testedOnly test for ABO required if no clinically significant antibodies currently or in history -Electronic (computer) a. If validated system and other requirements met b. Two determinations of ABO group, one on current sample c. Donor confirmed for ABo and Rh (on negative units) d. System can verify correct data entry and contains logic to alert if mistakes made102
992132907Antigen typing-Patients with clinically significant antibodies should receive antigen negative units - Probability of finding antigen negative units a. Multiply antigen negative (compatible) % converted to decimal b. C = (70% pos : 30% neg) ; JKa = (75% pos: 25% neg) .30 X .25 = .075 or 7.5% - probably find 8 units/100neg for C and Jka c. Only need three units - 8/100 = 3x, X =300/8 = 37.5 d. Need to screen 38 units to find 3 units C neg and JKa neg -Confirm antigen negative status by reacting cells with commercial preparations of antibody - QC rarely used antisera on day use a. Pos control- Heterozygous cell (anti-K tested with a Kk cell rather than a KK cell) b. Neg control - cell withough antigen (anti-K tested with a kk cell)103
992140958ABO identical is not available for RBCs- Decide what antibody (ies) are in the patients plasma - Transfused cells must lack corresponding antigens - O rbcs can be given to any alternative blood group- A, B or AB - AB can receive A, B and O rbcs - D positive can receive D positive or D negative: D negative however should only revieve D positive in emergency situation and D neg is not available. ( follow iwth RhIg if possible)104
992140964ABO identical is not available for plasma- Decide what antigen (s) are on the patients RBCs - Transfused plasma should lack the corresponding antibody (ies) - AB plasma can be given to any alternative blood group - A, B or O - Group O can receive A, B or AB plasma105
992340132Neonatal crossmatch- Initial ABO (cell type only) and D typing of infant RBCs and screen for unexpected antibodies using mother or infant serum or plasma -Crossmatch not necessary as long as antibody screen negative - No repeat testing is required for infants < 4 months if first two are met - To issue non- O RBCs, Not ABO compatible with maternal ABO,must test for passively acquired maternal ABO antibodies using antiglobulin phase - If clinically significant antibodies exist, infant must get antigen negative blood or units crossmatch compatible by antiglobulin crossmatch until antibody no longer detected - Infants <1200 g at birth, recommend units with reduced risk of CMV: recommend irradiated cells.106
992384429Intravascular hemolytic transfusion reaction-Transfused RBCs react with preformed antibodies in the recipient as transfusion is occurring - Usually due to clerical error involving ABO system - Fever- Most common symptom accompanied by chills, low back pain, anxiety - Physiological events a. Hemoglobinemia b. Hemoglobinuria c. Hyperbilirubinemia d. Can result in kidney failure and death107
992384430Extravascular hemolytic transfusion reaction- Usually due to anamnestic response to clinically significant antibodies such as Rh, Kell, Kidd and Duffy - Usually occurs after transfusion completed - Delayed transfusion reactions a. Hours to days after transfusion b. Indicated by NO rise or a ↓ in hemoglobin after transfusion c. positive DAT (key characteristic) d. Often due to Kidd antibodies108
992384431uticarial- Symptoms - itching and hives - If urticaria only symptoms, give antihistamine and transfusion may continue - Caused by donor antibodies to soluble plasma antigens109
992384432Febrile nonhemolytic (FNH)- Temperature rise associated with transfusion - Due to : Recipient preformed antibodies reacting with donor lymphocytes, granulocytes or platelets. Infusion of cytokines in donor bag from storage Prevention : Leukocyte-reduced blood components, Pre-storage leukoreduction prevents cytokine buildup and prevention usually considered after 2 separate febrile reactions110
992384433Allergic reaction- Recipient preformed IgE antibodies to soluble substance in plasma - Mild - uticarial - hives with itching - Give antihistamines and continue transfusion - Severe- anaphylaxis- systemic symptoms including hypotension, shock, sometimes death - Classic anaphylaxis- IgA deficient patient with anti-IgA reacting with IgA in donor plasma - Give washed cells or plasma components from IgA deficient donors111
992460463Transfusion- related acute lung injury- Acute respiratory insufficiency and bilateral pulmonary edema by X-ray without cardiac failure - includes chills, fever, and hypotension- TRALI - Donor antibodies to recipient HLA or neutrophil antigens - rarely, recipient antibodies to transfused granulocytes112
992460464Transfusion transmitted infections- Bacterial contamination is now most common since current tests detect most viruses. - All platelets must be tested for bacterial contamination before issue - Other diseases- HBV, HCV, HIV, HTLV, CMV, EBV, Babesiosis, Malaria, Chagas disease, West nile virus - "look back" - Identification of individuals who have recieved seronegative or untested blood from a donor later found to be infected - Must have mechanism to encourage reporting of possible transfusion- associated infections113
992460465Transfusion reaction workup- Clerical check- Most hemolytic transfusion reactions result from administering blood to the incorrect patient - Visual hemolysis- Compare with pre-transfusion sample- - DAT - May be negative if all incompatible cells are destroyed - ABO antibodies rapidly activate complement leading to lysis - ABO- ABO on post sample - Other tests - If error in any of above test :perform antibody screen pre/post-sample and re-crossmatch pre-post samples114
992460466Positive hemolysis/Negative DAT-Patient in sickle cell crisis - Thalassemia or G6PD deficient patient - Unit overheated/frozen - All cells hemolyzed115
992534767Hemolytic disease of the fetus and newborn etiology- Infant inherits antigen form biological father - Mother has corresponding IgG antibody (sensitized by previous pregnancies or transfusions) - Maternal antibody crosses placenta and coats fetal cells - Coated cells removed from fetal circulation causing anemia and hyperbilirubinemia - Bilirubin has affinity for lipid rich layers of skin and brain and is a ptoent neurotoxin causing brain damage (kernicterus) - Phottherapy is treatment of choice; exchange transfusion used when phototherapy fails116
992534768Intrauterine tranfusion- Supplies antigen negative blood - Unit selected a. Group O, D neg b. Negative for antigen to which maternal antibody directed (compatible with maternal antibody) c. Must be irradiated d. Should be from CMV seronegative donor or a leukoreduced unit if mother status CMV negative or unknown e. Should be negative for Hgb S f. Should be fresh- usually less than 7 days old117
992534769Exchange transfusion- Reduces bilirubin levels and removes maternal antibodies - Adds nitrogen negative cells and removes antibody-coated cells which would ↑ bilirubin levels when destroyed - Acceptable samples for crossmatch a. Maternal sample b. Eluate from infant's cells c. Infant serum Unit selection a. Negative for antigen to which maternal antibody directed (compatible with maternal antibody) b. Group ), if ABO HDN- D negative if Rh HDN c. Unit should be less than 5-7 days old collected in CPDA-1 d. Should be negative for Hbg S e. Should be irradiated118
992593972RH immune GlobinConcentrated Anti-D - Antepartum administration given at 28 weeks to all D negative women and again within 72 hours of delivery to D negative women with D positive infants119
992593984Preparation of RhIgIntramuscular RhIg -1 vial (300mg or 1500IU ) neutralizes 30 mL whole hemorrhage (FMH) or 15mL of RBCs IM or Intravenous preparation - 1 vial neutralizes 17 mL or 15mL RBCs120
992593985Rosette test-Used to screen if more than one vial of RhIg is needed - If positive - Kleihauer-Betke acid elution or flow cytometry will quantitate fetal maternal bleed (fetal cells resist acid elution and appear pink while adult cells are ghost cells)121
992593986Calculation of number of vials with addition of a safety vial1. Using Kleihauer-Betke test, count a total of 2000 cells, not the number of fetal and maternal cells 2. Divide the number of fetal cells by 2000 and multiple by 5000 to determine volume of fetal whole blood bleed a. 8 cells/2000 cells X 5000 mL = 20 mL fetal whole blood bleed 3. Divide by 30 since 1 vial of RhIg will neutralize 30mL of fetal blood a. 20mL/30mL= .66 vial 4. If decimal greater than .5 round up 1 and add 1 for safety factor 5. If decimal less that .5 round down and add 1 for safety factor 6.in this case, round up to .66 to 1 and add 1 for a total of 2 vials122
992593987RhIg can be given to a D negative recipient who receives D pos blood- Number of vials is determined by dividing the volume transfused by 30 for whole blood or 15 for RBCs - Should be considered for any women of childbearing age123
992593988HLA system- Human Leukocyte antigens - Cell surface glycoproteins a. Class 1 - On platelets and nucleated cells (mature RBCs may have small amounts) b. Class 2 - On B lymphs - Monocte/Macrophages, T lymphs and dendritic cells - Genes located on short arm of chromosome 6 (major histocompatibility complex)124
992593989HLA recognition- Contributes to self/non-self recongition - immune response - coordination of cellular and humoral responses - Second in importance only to ABO for long-term survival of transplanted solid organs and most important in hematopoietic progenitor cell transplantation125
992593990HLA function- Plays a role in a. Immune-mediated platelet refractoriness b. FNH-TR c. TRALI d. Posttransfusion graft-vs-host disease (GVHD) - Used for a. Susceptibility to certian disease b. Relationship (percentage) testing c. Forensic investigations126
992593991Detection HLADNA-based assay - High sensitivity and specificity Microlymphocytotoxicity - HLA-A, -B, -C, -DR, -DQ -Principle similiar to RBC typing only uses WBCs, rabbit complement, and dye exclusion technique to determine cell viability/death127
992593992Qc reagent- Reagents - each day of use a. Antihuman globulin b. Blood grouping reagents c. Antibody screening and reverse grouping cells Results - compare with previous results - inactivity implies reagent deterioration Antiglobulin reagent a. Required that IgG sensitized cells be added to all negative antiglobulin tests in antibody detection and compatibility testing b. negative test implies- Insufficient removal of serum proteins prior to addition of AHG (insufficient washing which allowed AHG to be neutralized by remianing serum). Ommision of AHG from procedure and must repeat tests128
992593993equipment QC- Hot blocks/waterbaths- 37°C ± 2 - observe temp day of use - Refrigerates - 1-6°C (as well as alarm activation checks) - Serofuge/cell washer - Timer checks, speed, Function- are cell buttons clearly delineated, is supernatant clear, do cells re-suspend with gentle agitation. -Component centrifuge - Thermometers- Tested initially129

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