Case Study WR Patient: WR, 93 y/o female No blood bank history Forward Typing Rh Typing Reverse Typing Anti-A Anti-B Anti-A,B Anti-D A 1 Cells B Cells

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1 Case Study WR Patient: WR, 93 y/o female No blood bank history Forward Typing Rh Typing Reverse Typing Anti-A Anti-B Anti-A,B Anti-D A 1 Cells B Cells What is the appropriate next step when investigating any ABO discrepancy? 1

2 Case Study WR Forward Typing Rh Typing Reverse Typing Anti-A Anti-B Anti-A,B Anti-D A1 Cells B Cells RT Incubation 0 2+ Select the type to investigate Reverse typing Possible Resolution: Increase incubation time Perform test at IAT Decrease incubation temperature 15 RT incubation Antibody screen control Screening Cells 15 RT I 0 II 0 III 0 Auto 0 2

3 Case Study WR Conclusion A Positive Group O and auto control non reactive Patient 93 y/o Weak Anti-B in plasma Extended incubation enhances antibody uptake 3

4 Case Study AE 36 year old female Diagnosis: Pregnancy #3 Medications: Pre-natal vitamins Transfusion History: No transfusions 4

5 Case Study AE Forward Type Rh Type Reverse Type Anti-A Anti-B Anti-D D Ctrl A 1 cells B cells Patient Screening Cell IS PEG/IAT I 0 0 II 0 0 III 0 0 5

6 Case Study AE Forward Type Rh Type Reverse Type Anti-A Anti-B Anti-A,B Anti-D D Ctrl A 1 cells A 2 cells B cells Patient RT Screening cell RT 30 I 0 II 0 Auto 0 6

7 Adsorption Testing Remove autoantibody from serum (warm/cold autoantibody) Separate antibody specificities (anti-kp b from anti-c) Confirm presence of an antigen ( weak A or B subgroup) 7

8 Adsorption Mechanics Human anti-a Patient Cells containing A antigen Combine and Incubate Cells coated with antibody Antigen/Antibody Complexes 8

9 Case Study AE Elution Washed cells coated with antibody Eluate containing antibody 9

10 Case Study AE Eluate Testing A cells A cells O cells Test Eluate Test Last Wash Control Eluate (A cells) Control LW Control Eluate (O cells) Control LW

11 Case Study AE Conclusion A Positive Patient Weak A antigen expression Adsorption/Elution confirms antigen presence 11

12 Case Study TC Patient: TC, 71 y/o male Knee replacement surgery Hemoglobin: 12.3 g/dl Medications: Acetaminophen Transfusion History: Pt reports he received some blood after a motor vehicle accident many years ago Physician ordered 2 units prbcs 12

13 Case Study TC ABO/Rh Forward Typing Anti-A Anti-B 4+ 0 Interpretation A Positive Rh Typing Anti-D 3+ Reverse Typing A 1 Cells B Cells

14 Case Study TC Antibody Screen Blood Group System Rh Kell Duffy Kidd Lewis P MNSs Lutheran D C c E e K k Fy a Fy b Jk a Jk b Le a Le b P 1 M N S s Lu a Lu b IS PEG IAT I R1R II R2R III rr Patient Cells

15 Case Study TC Crossmatches Anti-A,B IS PEG/IAT Donor Donor What are possible reasons for the incompatibility? Antibody to a low frequency antigen Donor unit exhibits a positive Direct Antiglobulin Test An ABO incompatible unit was selected 15

16 Case Study TC Donor 2: DAT Anti-IgG 1+ 16

17 Case Study TC Additional Crossmatch Anti-A Anti-B IS PEG/IAT Donor Conclusion ABO/RH A positive No antibodies identified Crossmatch compatible units ready for transfusion 17

18 Case Study PO 75 year old female Pre-op total knee replacement Current meds: Coumadin, vitamins No known transfusion history Order: type and hold for 2 units 18

19 Case Study PO Front Type Rh Reverse Type Interp. Anti-A Anti-B Anti-D A 1 cells B Cells A Positive Blood Group System Rh MNSs P Lewis Kell Duffy Kidd Lutheran D C E c e C w M N S s P 1 Le a Le b K k Fy a Fy b Jk a Jk b Lu a Lu b IS PEG/IAT 1 R 1 R R 2 R rr Auto

20 Selected Cell Testing Case Study PO Blood Group System Rh MNSs P Lewis Kell Duffy Kidd Lutheran D C E c e C w M N S s P 1 Le a Le b K k Fy a Fy b Jk a Jk b Lu a Lu b PEG/IAT 1 R 1 R R 2 R R 1 R

21 Case Study PO Unit Number IS PEG/AHG Unit ABO Anti-A Anti-B Anti-D Interp. Special Requirements Interpretation 12RE A NA Incompatible 55EE A NA Compatible 21

22 Case Study PO Blood Group System Rh MNSs P Lewis Kell Duffy Kidd Lutheran D C E c e C w M N S s P 1 Le a Le b K k Fy a Fy b Jk a Jk b Lu a Lu b 1 R 1 R R 1 R R 1 R R 2 R R 2 R r r r r rr rr rr PEG/IAT 22

23 Case Study PO Rh MNSs P Lewis Kell Duffy Kidd Lutheran D C E c e C w M N S s P 1 Le a Le b K k Fy a Fy b Jk a Jk b Lu a Lu b PEG/IAT 1 R 1 R R 2 R r r R 1 R R 1 R R 2 R r r rr rr rr

24 Case Study PO Blood Group System Rh MNSs P Lewis Kell Duffy Kidd Lutheran D C E c e C w M N S s P 1 Le a Le b K k Fy a Fy b Jk a Jk b Lu a Lu b 1 R 1 R Ficin/37 Ficin/IAT 2 R 1 R R 1 R R 2 R R 2 R r r r r rr rr rr Auto

25 Case Study PO Resolution: A Positive Anti-P1 at Ficin 37C/IAT and PEG/IAT 25

26 Case Study SY Patient: 60 y/o female Diagnosis: Gastrointestinal bleeding Medications: Not available No record of patient at hospital 26

27 Case Study SY ABO/Rh Forward Typing Anti-A Anti-B 0 4+ Interpretation B Positive Rh Typing Anti-D 3+ Reverse Typing A 1 Cells B Cells

28 Case Study SY Antibody Screen Blood Group System Rh Kell Duffy Kidd Lewis P MNSs Lutheran D C c E e K k Fy a Fy b Jk a Jk b Le a Le b P 1 M N S s Lu a Lu b IS Gel IAT I R1R II R2R III rr Patient Cells 0 0 Interpretation Positive 28

29 Case Study SY Blood Group System Rh Kell Duffy Kidd Lewis P MNSs Lutheran D C c E e K k Fy a Fy b Jk a Jk b Le a Le b P 1 M N S s Lu a Lu b Gel IAT 1 R1R R1R w+ 3 R2R R2R rr w+ 6 rr What testing should be performed to identify the reactivity? 29

30 Case Study SY Phenotype Autocontrol: Negative No record of transfusion Critical to watch for mixed field C c E e K k Fy a Fy b Jk a Jk b M N S s

31 Case Study SY Patient is negative for E, K, Fy a, M, and S Test two phenotypically similar cells Reactive High incidence antibody Non-reactive Multiple antibodies Blood Group System Rh Kell Duffy Kidd Lewis P MNSs Lutheran D C c E e K k Fy a Fy b Jk a Jk b Le a Le b P 1 M N S s Lu a Lu b Gel IAT 1 R1R rr

32 Case Study SY Blood Group Systemr Rh Kell Duffy Kidd Lewis P MNSs Lutheran D C c E e K k Fy a Fy b Jk a Jk b Le a Le b P 1 M N S s Lu a Lu b GEL IAT 1 R2R RzR2 + w rr rr rr rr rr rr rr rr

33 Case Study SY Conclusion ABO/RH B Positive Antibody ID Anti-Fy a at PEG/IAT Anti-S at PEG/IAT Transfusion Recommendations Transfuse B or O positive units negative for Fy a and S antigens 33

34 Case Study GI 56 year old female GI Bleed Current meds: None Transfusion History: Transfused 2 days ago Blood Bank History A positive 34

35 Case Study GI Front Type Rh Reverse Type Interp. Anti-A Anti-B Anti-D A 1 cells B Cells A Positive Blood Group System Rh MNSs P1 Lewis Kell Duffy Kidd Lutheran D C E c e M N S s P1 Le a Le b K k Fy a Fy b Jk a Jk b Lu a Lu b IS PEG/IAT 1 R 1 R R 2 R Auto

36 Case Study GI Poly IgG C3d Control Positive IgG DAT 2+mf 2+mf

37 Case Study GI 37

38 Elution Indications Investigation of a positive DAT result Autoantibodies Recently transfused patients Hemolytic Disease of the Fetus and Newborn (HDFN)

39 Case Study GI Rh MNSs Lewis P Kell Duffy Kidd Lutheran D C E c e M N S s Le a Le b P1 K k Fy a Fy b Jk a Jk b Lu a Lu b Eluate/PEG/IAT Last Wash w

40 Questions? 40

41 References Nedelcu E Et al. Transfusion 2017;57(S3):148A-149A. Abstract no. CP218. Velliquette RW et al. Transfusion 2017;57(S3):159A. Abstract no. CP ation&rlz=1c1ggrv_enus794us797&tbm=isch&sourc e=iu&ictx=1&fir=keuxgorlqbu9rm%253a%252c7pb2 V5nz_Xp9SM%252C_&usg= -J3EK- JyP41JB6dQfxNj8aymbjY%3D&sa=X&ved=0ahUKEwipl ZnW3vHbAhVNaFAKHR-lCxIQ9QEIXDAH#imgrc=_ 41

Chin J Blood Transfusion Nov 2013 Vol. 26 No LIMS electronic issue EI and Healthcare Products Regulatory Agency MHRA 2. 1

2013 11 26 11 Chin J Blood Transfusion Nov 2013 Vol. 26 No. 11 郭永建 1161 BCSH 英国血液学标准委员会 2012 年赵国华赵维齐 100021 BCSH R457. 1 A 1004-549X 2013 11-1161-12 British Committee for Standards in Haematology BCSH