An accurate genetic assay to identify human neutrophil antigen 2 deficiency.
An accurate genetic assay to identify human neutrophil antigen 2 deficiency.
复制标题
用于识别人类中性粒细胞抗原 2 缺陷的准确基因检测。
DOI：
10.1111/tme.12936
复制
发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Wu,Jianming
中科院分区：
文献类型：
--
作者：
Li,Yunfang;Schuller,RandyM;Wu,Jianming
HNA-2是一种重要的中性粒细胞抗原，与多种人类疾病密切相关。HNA-2缺陷或HNA-2 null是在3%-5%的美国人中观察到的常见表型。HNA-2缺失个体有产生同种抗体（或同种抗体）的风险，这些抗体在输血相关急性肺损伤、免疫性中性粒细胞减少症和骨髓移植失败中起重要作用。我们先前证明了CD 177编码SNP 787 A> T（c.787A > T）是HNA-2缺陷最重要的遗传决定因素。研究设计和方法采用一种新的聚合酶链反应（PCR）策略来确定CD 177 SNP c.787A > T的基因型。在简化的PCR检测中，所有等位基因特异性引物和内部对照引物都包括在同一反应中，这确保了检测的可靠性。结果396例HNA-2缺陷症患者CD 177 SNP c787 A> T基因型与巢式TaqMan法、桑格测序法检测结果的一致性均为100%。在396例受试者中，18例CD 177 STP纯合基因型供者均为HNA-2 null.ConclusionThe novel PCR-based genotyping assay is accurate to identify HNA-2 deficiency individuals and is suitable for clinical laboratory.此外，创新的高通量巢式TaqMan检测将用于大规模人群筛查和HNA-2缺陷的遗传研究。
ObjectiveWe aimed to develop accurate and user‐friendly genetic assays to identify the inherited neutrophil antigen‐2 (HNA‐2) deficiency in humans.BackgroundHNA‐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 theCD177coding 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 MethodsA novel polymerase chain reaction (PCR) strategy was used to determine genotypes of theCD177SNP 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.ResultsCD177SNP 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 theCD177STP homozygous genotype were HNA‐2 null.ConclusionThe 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.