28
Robert Stamps, RSM, RCI Sheffield
Robert Stamps, RSM, RCI Sheffield
Robert Stamps, RSM, RCI Sheffield
ABO blood groups - Landsteiner 1900Rh blood groups Levine and Stetson 1939 Landsteiner and Weiner 1940AHG anti-human globulin Coombs 1945LISS Low ionic strength solution 1970 s Monoclonal blood typing sera 1980 s Gel column technology - Invented1985 DiaMedintroduced test system in 1988
Robert Stamps, RSM, RCI Sheffield
Anti-globulin reagent - AHG
Robert Stamps, RSM, RCI Sheffield
Higher risk of antibody production in multi-transfused patientsEthnic minority patients at greater riskUp to 30% become alloimmunisedMost produce Rh (anti-D, C, E, c, e) and K antibodies
Robert Stamps, RSM, RCI Sheffield
Spanos et al, Red cell alloantibodies in patients with thalassemia, Vox Sang 1990; 58Rh 34%, K 30%, MSs 8%, Jk 8%, Fy 4%Ameen et al, RBC allo and auto immunisation among transfusion-dependent Arab thalassemiapatients, Transfusion 2003;43Rh 49%, K 31%, Jk 5%, Fy 3%11% developed autoantibodies -immunomodulationAnti-E was most commonly detected antibody in both studies
Robert Stamps, RSM, RCI Sheffield
Delayed haemolytic transfusion reaction
45 year old female, hysterectomy O PosTransfused 2 units3 days later jaundice, raised LDH, Hb 8.5DAT Pre transfusion NegativeDAT Post transfusion Positive C3d and weak IgGAntibody screen Pre repeat neg, post -Pos
Robert Stamps, RSM, RCI Sheffield
DAT Positive IgGDAT Positive IgG + C3d
Robert Stamps, RSM, RCI Sheffield
Panel results
ID Panel NBS REAGENTS
Lot R1412016 NAME NUT Mary DATE OF BIRTH 25/12/1964 GROUP O Pos R1r Exp. 31/12/2501 HOSPITAL NCH HOSPITAL No. SAMPLE No. 1
Rh CW
C c D E e M
N S s P1 Lua
K k Kpa
Lea
Leb
Fya
Fyb
Jka
Jkb
Other IAT Enz IAT
1 R1WR1 + +
0
+
0
+
+
+
0
+
0 0 0 + 0 + 0 + 0 + 0
2 4 2 R1
R1 0 +
0
+
0
+
+
0
+
0
2+ + + + 0 0 0 0 + 0 +
0 0 3 R2 R2 0 0
+
+
+
0
+
+
0
+
0 0 0 + 0 0 + 0 + 0 +
0 0 4 r ` r 0 +
+
0
0
+
+
+
0
+
4+ 0 0 + 0 0 0 0 + 0 + Yk (a-) 0 0 5 r``r 0 0
+
0
+
+
+
+
+
0
5+ 0 0 + 0 0 + 0 + + 0
2 4 6 r r 0 0
+
0
0
+
0
+
+
0
0 0 0 + 0 0 + + + 0 +
0 0 7 r r 0 0
+
0
0
+
+
+
+
+
4+ 0 + + 0 + 0 + 0 + 0
2 4 8 r r 0 0
+
0
0
+
0
+
0
+
2+ + 0 + 0 0 + + 0 0 +
0 0 9 r r 0 0
+
0
0
+
0
+
0
+
5+ + 0 + + 0 + 0 + + 0
2 4 10 r r 0 0
+
0
0
+
+
0
0
+
4+ 0 + 0 0 0 + 0 + 0 +
0 0
Auto
2
Robert Stamps, RSM, RCI Sheffield
Antibody level may be too low to detectAfter transfusion rapid increase in antibody concentration after 3-7 daysMay be rapid destruction of transfused cellsAnti-Jka most commonly describedCan occasionally be seen after ABO incompatible transfusion in elderly or immune suppressed
Robert Stamps, RSM, RCI Sheffield
Female patient reacted to one of three units (rigors and vomiting)Standard panel and repeat antibody screen negativeUnit incompatible by IATAntibody to low incidence antigen Identified as anti-Vw MNS systemAntigen very rare in UK (0.06%) Antibody present in about 1%
Robert Stamps, RSM, RCI Sheffield
Serological Investigation of Transfusion Reaction
Pre-transfusion sample - ABO and Rhphenotype - repeat red cell antibody screen and crossmatch. DATPost transfusion sample - ABO Rh D type -antibody investigation and repeat crossmatch. DATEluate on post transfusion sample Can be useful even when DAT negative
Robert Stamps, RSM, RCI Sheffield
Removal of immunoglobulin (usually IgG) from patient/donor red cells Allows serological investigation of bound antibody to determine nature and specificityMany methods available Heat Landsteiner-Miller Solvents Rubin s Etheracid solution pH 3.0
Robert Stamps, RSM, RCI Sheffield
Jaundiced baby with Positive DAT
2 day old baby with low Hb and jaundiceMaternal antibody screen NegativeDAT on Baby Positive IgGMother group O baby group A
Robert Stamps, RSM, RCI Sheffield
Maternal serum contains IgG anti-A as well as IgM anti-AIgG anti-A can cross placentaBabies group A antigens strengthen after birthIf exchange transfusion required Group O blood cross-matched x maternal plasma
Robert Stamps, RSM, RCI Sheffield
Antenatal patient - Panel results
ID Panel NBS REAGENTS
Lot R1412016 NAME Sickely Pat DATE OF BIRTH 18/12/1984 GROUP B Neg rr Exp. 31/12/2501 HOSPITAL NCH HOSPITAL No. SAMPLE No. 1
Rh CW
C
c D
E e M
N S s P1 Lua
K k Kpa
Lea
Leb
Fya
Fyb
Jka
Jkb
Other IAT Enz
1 R1WR1 + +
0
+
0
+
+
+
0
+
0 0 0 + 0 + 0 + 0 + 0
3 5 2 R1
R1 0 +
0
+
0
+
+
0
+
0
2+ + + + 0 0 0 0 + 0 +
3 5 3 R2 R2 0 0
+
+
+
0
+
+
0
+
0 0 0 + 0 0 + 0 + 0 +
3 5 4 r ` r 0 +
+
0
0
+
+
+
0
+
4+ 0 0 + 0 0 0 0 + 0 + Yk (a-) 0 0 5 r``r 0 0
+
0
+
+
+
+
+
0
5+ 0 0 + 0 0 + 0 + + 0
0 0 6 r r 0 0
+
0
0
+
0
+
+
0
0 0 0 + 0 0 + + + 0 +
0 0 7 r r 0 0
+
0
0
+
+
+
+
+
4+ 0 + + 0 + 0 + 0 + 0
0 0 8 r r 0 0
+
0
0
+
0
+
0
+
2+ + 0 + 0 0 + + 0 0 +
0 0 9 r r 0 0
+
0
0
+
0
+
0
+
5+ + 0 + + 0 + 0 + + 0
0 0 10 r r 0 0
+
0
0
+
+
0
0
+
4+ 0 + 0 0 0 + 0 + 0 +
0 0 Auto
0
Robert Stamps, RSM, RCI Sheffield
Patient 32 weeks pregnant Attended A/E with PV bleedPanel revealed anti-DIs anti-D due to 28 week prophylactic anti-D injection?Quantitate anti-D and check history if injected and level < 0.15iu likely to be remains of injectionImportant not to assume it is prophylactic
Robert Stamps, RSM, RCI Sheffield
Myelodysplastic syndromesThalassaemiasSickle cell diseaseSevere aplastic anaemiasWarm/cold autoimmune haemolytic anaemiasOther congenital or acquired chronic anaemias
Robert Stamps, RSM, RCI Sheffield
Most common phenotype is Ror (cDe/cde) - 56%68% Fya-b-Only rarely form Fy antibodiesUsually anti-Fya - although Fyb not present on red cells it is present on other tissuesMay eventually form anti-Fy3Supply of blood more difficult if patient not Ror ieR1R1 or RhD-
Robert Stamps, RSM, RCI Sheffield
RhD- (Cde/cde) femaleMultitransfused D- bloodAlloantibodies present Anti-E, K, M, S, Fya, Jkb
Lea+b, KnopsHyperhaemolysis event after ignoring anti-MEventually formed anti-Fy3Very few O D-, M-, S-, K-, Fya-b-, Jkb- donors
Robert Stamps, RSM, RCI Sheffield
Group O R1R1 patient with anti-cDeveloped anti-Fy3Only TWO suitable donors in UKBlood sourced from Amsterdam
Robert Stamps, RSM, RCI Sheffield
HTLA High titre low avidityCommon in multi-transfused patientsMany specificities Ch, Rg, Yka, Yta, Sla, Kna etcAll clinically insignificant but may mask significant alloantibodiesPhenotyped matched blood
Robert Stamps, RSM, RCI Sheffield
000Auto
4550+0++000+05+0+0rr10
455+0+0+0++004+0++rr9
455++0+0+0+0+30+0+rr8
455+0+00+0++03+0++rr7
4550++0+00+0050++0rr6
455+00++00+003++0+r r5
455+0+0+00+000+0+0r r4
4550++0+00+003+0+0R2R23
4550+0++00++040+0+R1R12
455+0++0+0+000+0+0R1wR11
IATEnzSAL R/T
JkbJkaFybFyaLebLeaKp
akKLuaP1sSNMRh
Antibody identification panel
5555555055000
Cell 3Cell 2Cell 1O R1rB rrA2 rrA1 rr
Antibody Screen (IAT)Reverse Group (cells)Reagent ControlAnti-D (2)Anti-D (1)Anti-A,BAnti-BAnti-A
Blood grouping
Robert Stamps, RSM, RCI Sheffield
Over 50% samples referred to reference lab have autoantibodiesMany require regular transfusionsMost are warm type IgG autoantibodiesAdsorption of autoantibody required to detect underlying alloantibodies Autoadsorption or allogeneic adsorptionCrossmatch is positive and blood issued as suitable
Robert Stamps, RSM, RCI Sheffield
Autoadsorbed patient typical serologyID Panel NBS REAGENTS
Lot R1412016 NAME Hill Harry DATE OF BIRTH 25/01/1926 GROUP O Pos R2r Exp. 31/12/2501 HOSPITAL NCH HOSPITAL No. SAMPLE No. 1
Rh CW
C
c D
E e M
N
S s P1 Lua
K k Kpa
Lea
Leb
Fya
Fyb
Jka
Jkb
Other IAT Enz IAT
Auto abs xIAT
1 R1WR1 + +
0
+
0
+
+
+
0
+
0 0 0 + 0 + 0 + 0 + 0
4 5 3 2 R1
R1 0 +
0
+
0
+
+
0
+
0
2+ + + + 0 0 0 0 + 0 +
4 5 0 3 R2 R2 0 0
+
+
+
0
+
+
0
+
0 0 0 + 0 0 + 0 + 0 +
4 5 0 4 r ` r 0 +
+
0
0
+
+
+
0
+
4+ 0 0 + 0 0 0 0 + 0 + Yk (a-) 4 5 0 5 r``r 0 0
+
0
+
+
+
+
+
0
5+ 0 0 + 0 0 + 0 + + 0
4 5 0 6 r r 0 0
+
0
0
+
0
+
+
0
0 0 0 + 0 0 + + + 0 +
4 5 2 7 r r 0 0
+
0
0
+
+
+
+
+
4+ 0 + + 0 + 0 + 0 + 0
4 5 3 8 r r 0 0
+
0
0
+
0
+
0
+
2+ + 0 + 0 0 + + 0 0 +
4 5 3 9 r r 0 0
+
0
0
+
0
+
0
+
5+ + 0 + + 0 + 0 + + 0
4 5 0 10 r r 0 0
+
0
0
+
+
0
0
+
4+ 0 + 0 0 0 + 0 + 0 +
4 5 0
Auto
5
Adsorption reveals anti-Fya
Robert Stamps, RSM, RCI Sheffield
Allogeneic adsorption typical results
ID Panel NBS REAGENTS
Lot R1412016 NAME GOK Jane DATE OF BIRTH 18/12/1929
GROUP O Pos R1R1
Exp. 31/12/2501 HOSPITAL NCH HOSPITAL No. SAMPLE No. 7
Rh CW C
c D
E e M
N S s P1 Lua K k Kpa Lea Leb Fya Fyb Jka Jkb Other IAT Enz
IAT Ads x rr
Ads x R1R1
1 R1WR1 + +
0
+
0
+
+
+
0
+
0 0 0 + 0 + 0 + 0 + 0
5 5 5 0 2 R1 R1 0 +
0
+
0
+
+
0
+
0
2+ + + + 0 0 0 0 + 0 +
5 5 5 0 3 R2 R2 0 0
+
+
+
0
+
+
0
+
0 0 0 + 0 0 + 0 + 0 +
5 5 5 4 4 r ` r 0 +
+
0
0
+
+
+
0
+
4+ 0 0 + 0 0 0 0 + 0 + Yk (a-) 5 5 0 2 5 r``r 0 0
+
0
+
+
+
+
+
0
5+ 0 0 + 0 0 + 0 + + 0
5 5 0 4 6 r r 0 0
+
0
0
+
0
+
+
0
0 0 0 + 0 0 + + + 0 +
5 5 0 4 7 r r 0 0
+
0
0
+
+
+
+
+
4+ 0 + + 0 + 0 + 0 + 0
5 5 0 4 8 r r 0 0
+
0
0
+
0
+
0
+
2+ + 0 + 0 0 + + 0 0 +
5 5 0 4 9 r r 0 0
+
0
0
+
0
+
0
+
5+ + 0 + + 0 + 0 + + 0
5 5 0 4 10 r r 0 0
+
0
0
+
+
0
0
+
4+ 0 + 0 0 0 + 0 + 0 +
5 5 0 4
5
Adsorption reveals allo anti-c and auto anti-D
Robert Stamps, RSM, RCI Sheffield
CHAD IgM wide thermal range cold auto anti-IAdsorption not usually requiredPre-warming of tests usually successfulRabbit stroma useful if adsorption required
Robert Stamps, RSM, RCI Sheffield
All donations are tested for Rh (C,c,E,e) and KA number of units are typed for (related to demand and usually rr, R1R1 and R2R2) M,S,Cw,Jka,Jkb,Fya,Fyb and HbSVery rare units offered to frozen blood bankRCI inform Donor Testing of any problematic patientsTesting ensure suitable units are held for named patients
Robert Stamps, RSM, RCI Sheffield
Red cell genotyping using DNAShirey RS et al, Prophylactic antigen matched donor blood for patients with warm autoantibodies, an algorithm for transfusion management, Transfusion 2002, vol 42
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