Sudan Tao, Xuan You, Jielin Wang, Wei Zhang, Ji He, Faming Zhu
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引用次数: 0
Abstract
Killer cell immunoglobulin-like receptor (KIR) and human leukocyte antigen (HLA) play crucial roles in regulating NK cell activity. Here, we report a real-time quantitative PCR (qPCR) to genotype all KIR genes and their copy numbers simultaneously. With 18 pairs of locus-specific primers, we identified KIR genes by Ct values and determined KIR copy number using the 2-∆Ct method. Haplotypes were assigned based on KIR gene copy numbers. The real-time qPCR results were consistent with the NGS method, except for one sample with KIR2DL5 discrepancy. qPCR is a multiplex method that can identify KIR copy number, which helps obtain a relatively accurate haplotype structure, facilitating increased KIR research in laboratories where NGS or other high-resolution methods are not available.
杀伤细胞免疫球蛋白样受体（KIR）和人类白细胞抗原（HLA）在调节 NK 细胞活性方面起着至关重要的作用。在此，我们报告了一种实时定量 PCR（qPCR）方法，可同时对所有 KIR 基因及其拷贝数进行基因分型。利用 18 对基因座特异性引物，我们通过 Ct 值确定了 KIR 基因，并用 2-∆Ct 法确定了 KIR 拷贝数。根据 KIR 基因拷贝数分配单倍型。实时 qPCR 的结果与 NGS 方法一致，只有一个样本的 KIR2DL5 存在差异。qPCR 是一种可识别 KIR 拷贝数的多重方法，有助于获得相对准确的单倍型结构，从而有助于在无法使用 NGS 或其他高分辨率方法的实验室开展更多的 KIR 研究。
期刊介绍：
Immunogenetics publishes original papers, brief communications, and reviews on research in the following areas: genetics and evolution of the immune system; genetic control of immune response and disease susceptibility; bioinformatics of the immune system; structure of immunologically important molecules; and immunogenetics of reproductive biology, tissue differentiation, and development.
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