An accurate genetic assay to identify human neutrophil antigen 2 deficiency

• Published in: Transfusion medicine (Oxford, England) Vol. 33; no. 1; pp. 68 - 74
• Main Authors: Li, Yunfang, Schuller, Randy M., Wu, Jianming
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.02.2023
• Subjects: CD177; genetic assay; Genotype; HNA‐2 deficiency; Homozygote; Humans; Isoantigens - genetics; Neutrophils; polymorphisms; Reproducibility of Results; genetic assay; polymorphisms; HNA-2 deficiency; CD177
• ISSN: 0958-7578; 1365-3148; 1365-3148
• DOI: 10.1111/tme.12936

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.