Evaluation of Diagnostic Tests by Evanescence Biosensor Technology for Rapid Phenotyping of the Human Platelet Alloantigens 1a and 5b.
Genotyping
HPA phenotyping
Neonatal alloimmune thrombocytopenia
Journal
Transfusion medicine and hemotherapy : offizielles Organ der Deutschen Gesellschaft fur Transfusionsmedizin und Immunhamatologie
ISSN: 1660-3796
Titre abrégé: Transfus Med Hemother
Pays: Switzerland
ID NLM: 101176417
Informations de publication
Date de publication:
Dec 2019
Dec 2019
Historique:
received:
26
07
2018
accepted:
07
09
2018
entrez:
15
1
2020
pubmed:
15
1
2020
medline:
15
1
2020
Statut:
ppublish
Résumé
The human platelet alloantigens (HPA) HPA-1a and HPA-5b are located on glycoproteins on the platelet surface and are the most relevant to cause neonatal alloimmune thrombocytopenia (NAIT). The antigens are defined by single nucleotide polymorphisms (SNPs) in the glycoprotein genes, and the antigen status can be determined by genotyping the SNPs. However, genotyping is time-consuming and costly depending on the method and sample throughput. Here, we tested the reliability of the evanescence wave based fluorescence (EVA) biosensor technology for the rapid phenotyping of the HPA-1a and HPA-5b antigens on blood donor samples in two laboratories. HPA-1a and HPA-5b phenotyping was performed on EDTA blood samples from 336 blood donors (Lyon: 216 donors; Mannheim: 120 donors) using EVA typing assays and the biosensor system (Davos Diagnostics, Davos, Switzerland). For genotyping, validated PCR-SSP and TaqMan-PCR methods were used. HPA-1a phenotyping was positive for all samples with HPA-1aa (n = 244; EVA value 807 ± 167 U/s) and HPA-1ab (n = 82; 542 ± 110 U/s) genotypes. All samples (n = 10) with negative EVA values (<10 U/s) had the HPA-1bb genotype. HPA-5b phenotyping was negative for all HPA-5aa genotypes (n = 267) and positive for the HPA-5ab (n = 66; 83 ± 22 U/s) and HPA-5bb (n = 3; 118 ± 25 U/s) genotypes. EVA values from heterozygotes were significantly lower compared to HPA-1a or HPA-5b homozygotes. A strong correlation of the EVA values with the platelet count in the blood samples was observed. EVA is a reliable method for rapid phenotyping of the clinically relevant HPA-1a and HPA-5b platelet antigens. All phenotyping results were 100% concordant with the HPA-1 or HPA-5 genotype. The test can be performed from only 10 µl of fresh or frozen blood samples within less than 15 min time-to-result.
Sections du résumé
BACKGROUND
BACKGROUND
The human platelet alloantigens (HPA) HPA-1a and HPA-5b are located on glycoproteins on the platelet surface and are the most relevant to cause neonatal alloimmune thrombocytopenia (NAIT). The antigens are defined by single nucleotide polymorphisms (SNPs) in the glycoprotein genes, and the antigen status can be determined by genotyping the SNPs. However, genotyping is time-consuming and costly depending on the method and sample throughput. Here, we tested the reliability of the evanescence wave based fluorescence (EVA) biosensor technology for the rapid phenotyping of the HPA-1a and HPA-5b antigens on blood donor samples in two laboratories.
METHODS
METHODS
HPA-1a and HPA-5b phenotyping was performed on EDTA blood samples from 336 blood donors (Lyon: 216 donors; Mannheim: 120 donors) using EVA typing assays and the biosensor system (Davos Diagnostics, Davos, Switzerland). For genotyping, validated PCR-SSP and TaqMan-PCR methods were used.
RESULTS
RESULTS
HPA-1a phenotyping was positive for all samples with HPA-1aa (n = 244; EVA value 807 ± 167 U/s) and HPA-1ab (n = 82; 542 ± 110 U/s) genotypes. All samples (n = 10) with negative EVA values (<10 U/s) had the HPA-1bb genotype. HPA-5b phenotyping was negative for all HPA-5aa genotypes (n = 267) and positive for the HPA-5ab (n = 66; 83 ± 22 U/s) and HPA-5bb (n = 3; 118 ± 25 U/s) genotypes. EVA values from heterozygotes were significantly lower compared to HPA-1a or HPA-5b homozygotes. A strong correlation of the EVA values with the platelet count in the blood samples was observed.
CONCLUSION
CONCLUSIONS
EVA is a reliable method for rapid phenotyping of the clinically relevant HPA-1a and HPA-5b platelet antigens. All phenotyping results were 100% concordant with the HPA-1 or HPA-5 genotype. The test can be performed from only 10 µl of fresh or frozen blood samples within less than 15 min time-to-result.
Identifiants
pubmed: 31933574
doi: 10.1159/000493556
pii: tmh-0046-0440
pmc: PMC6944947
doi:
Types de publication
Journal Article
Langues
eng
Pagination
440-445Informations de copyright
Copyright © 2018 by S. Karger GmbH, Freiburg.
Déclaration de conflit d'intérêts
Claudio Rhyner and Manfred Schawaller are employed by Davos Diagnostics AG, Switzerland. All other authors declare no conflict of interest. The EVA HPA-1a and HPA-5b chips required for this study were provided by Davos Diagnostics for free.
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