An accurate genetic assay to identify human neutrophil antigen 2 deficiency

Yunfang Li, Randy M. Schuller, Jianming Wu

Research output: Contribution to journalArticlepeer-review

Abstract

Objective: We aimed to develop accurate and user-friendly genetic assays to identify the inherited neutrophil antigen-2 (HNA-2) deficiency in humans. Background: HNA-2 is one of the most important neutrophil antigens implicated in a number of human disorders. HNA-2 deficiency or HNA-2 null is a common phenotype observed in 3%–5% Americans. HNA-2 null individuals are at risk to produce isoantibodies (or alloantibodies) that play important roles in transfusion-related acute lung injury, immune neutropenia, and bone marrow graft failure. We previously demonstrated that the CD177 coding SNP 787A > T (c.787A > T) is the most important genetic determinant for HNA-2 deficiency. However, reliable genetic assays are not available for routine clinical laboratory application up to now. Study Design and Methods: A novel polymerase chain reaction (PCR) strategy was used to determine genotypes of the CD177 SNP c.787A > T. In the simplified PCR assay, all allele specific primers and internal control primers were included in the same reaction, which ensures reliability of the assay. In addition, a novel high-throughput nested TaqMan assay was developed to determine genotypes of c.787A > T for large population genetic analysis of HNA-2 deficiency. Results: CD177 SNP c787A > T genotypes of 396 subjects were 100% concordant among the single PCR reaction method, the nested TaqMan assay, and Sanger Sequencing analysis. Out of 396 subjects, all 18 donors with the CD177 STP homozygous genotype were HNA-2 null. Conclusion: The novel PCR-based genotyping assay is accurate to identify HNA-2 deficient individuals and is suitable for clinical laboratories. In addition, the innovative high-throughput nested TaqMan assay will be useful for large-scale population screens and genetic studies of HNA-2 deficiency.

Original languageEnglish (US)
Pages (from-to)68-74
Number of pages7
JournalTransfusion Medicine
Volume33
Issue number1
DOIs
StatePublished - Feb 2023

Bibliographical note

Funding Information:
This study was supported by a grant from the National Institute of Health, United States (R21AI149395 to Jianming Wu). We greatly appreciated Memorial Blood Center in St. Paul for donor recruitment and sample collection.

Publisher Copyright:
© 2022 The Authors. Transfusion Medicine published by John Wiley & Sons Ltd on behalf of British Blood Transfusion Society.

Keywords

  • CD177
  • HNA-2 deficiency
  • genetic assay
  • polymorphisms

PubMed: MeSH publication types

  • Journal Article

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