A modified rapid monoclonal antibody-specific immobilization of platelet antigen assay for the detection of human platelet antigen (HPA) antibodies: a multicentre evaluation.
BACKGROUND
The monoclonal antibody-specific immobilization of platelet antigens (MAIPA) assay is the cornerstone technique for the detection and identification of human platelet antigen (HPA) antibodies. However, the original technique described by Kiefel and colleagues requires approximately 8 h adding to diagnostic delay. Moreover, proficiency exercises indicate that there are substantial variations in the MAIPA protocol, and that these may account for interlaboratory differences in sensitivity and specificity.


METHODS
A review of current MAIPA assay protocols from six laboratories together with performance in quality-assessment schemes identified several key variables potentially affecting the assay results. An optimized protocol was derived and assay time reduced to 5 h. The modified rapid MAIPA (MR-MAIPA) assay was evaluated using 61 samples with a range of HPA antibodies typically encountered in cases of fetomaternal alloimmune thrombocytopenia (n = 22), post-transfusion purpura (n = 8), platelet refractoriness (n = 7) and other platelet immune conditions (n = 24). The sensitivity of the assay was assessed using three international standards and the recombinant HPA-1a antibody CamTran007. The results obtained were compared with the original findings obtained with the local MAIPA assays. In addition, four different glycoprotein IIb/IIIa capture monoclonal antibodies were evaluated for their effect on assay sensitivity.


RESULTS
Complete concordance was found between the original MAIPA results and those obtained with the new assay when testing a selected panel of clinical samples. The modified assay had nanogram level sensitivity for the detection of HPA-1a antibodies and titration of HPA-1a and HPA-5b antibody sensitivity standards yielded end-points equal to or greater than the mean recorded in international workshops.


CONCLUSIONS
The MR-MAIPA assay offers improved turnaround for the detection of HPA antibodies without loss of sensitivity.
Campbell
K
K
National Health Service Blood and Transplant, Cambridge Centre, Long Road, Cambridge CB2 2PT, UK.
Rishi
K
K
Howkins
G
G
Gilby
D
D
Mushens
R
R
Ghevaert
C
C
Metcalfe
P
P
Ouwehand
W H
WH
Lucas
G
G
D000911:Antibodies, Monoclonal; D016824:Antigens, Human Platelet / Q000276:immunology*; D000094902:Blood Banking / Q000379:methods*; D006403:Hematologic Tests / Q000379:methods; D006801:Humans; D007118:Immunoassay / Q000379:methods; D007518:Isoantibodies / Q000032:analysis* / Q000097:blood; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D016824:Antigens, Human Platelet; D007518:Isoantibodies