A new bead-based human platelet antigen antibodies detection assay versus the monoclonal antibody immobilization of platelet antigens assay

Transfusion. 2014 Jun;54(6):1486-92. doi: 10.1111/trf.12509. Epub 2013 Dec 3.

Abstract

Background: The performance of a newly developed Luminex bead-based platelet (PLT) antibody detection method (PAKLx) was compared with the monoclonal antibody immobilization of PLT antigens (MAIPA) assay and the LifeScreen Deluxe Luminex bead-based HLA Class I antibody detection method (LMX).

Study design and methods: Six sera containing anti-human PLT antigen (HPA)-1a (n=2), HPA-1b, HPA-2b, HPA-3a, or HPA-5b were tested in titration. A total of 194 sera, including HPA-1a, -1b, -2a, -2b, -3a, -5a, and -5b antibodies with or without HLA antibodies (n=63); glycoprotein (GP) IV antibodies (n=1); PLT autoantibodies (n=3); HLA antibodies (n=45); and samples with no PLT-reactive antibodies (n=82), were tested in both assays.

Results: Comparable levels of sensitivity were obtained for the MAIPA and PAKLx. The PAKLx showed four (6%) false-negative results in 67 sera with HPA or GP-reactive antibodies: anti-HPA-3a (n=1) or anti-HPA-5b (n=3). The PAKLx showed in 10 of the total 194 samples (5%) the presence of antibodies not detected by the MAIPA. This concerned broadly GP-reactive antibodies (n=7), anti-GPIIb/IIIa combined with anti-HPA-3a (n=1), anti-HPA-1a (borderline, n=1), and anti-GPIV (n=1). Testing 175 sera for anti-HLA Class I antibodies in the PAKLx and LMX showed four discrepant results: PAKLx negative and LMX positive, n=3 and n=1, respectively.

Conclusion: For the vast majority of the specimens tested (93%) the results of the PAKLx were in concordance with the MAIPA. The PAKLx is a fast, easy to perform, and sensitive PLT antibody screening method.

MeSH terms

  • Antibodies / analysis*
  • Antibodies / immunology*
  • Antibodies, Monoclonal / analysis*
  • Antibodies, Monoclonal / immunology*
  • Antigens, Human Platelet / immunology*
  • Biological Assay / methods*
  • Humans
  • Integrin beta3

Substances

  • 2a alloantigen, human
  • 3a alloantigen, human
  • 5a alloantigen, human
  • 5b alloantigen, human
  • Antibodies
  • Antibodies, Monoclonal
  • Antigens, Human Platelet
  • ITGB3 protein, human
  • Integrin beta3
  • human platelet antigen 1b