ARL15、HLA-DMA基因多态性与中国西北地区汉族人群类风湿性关节炎易感性相关

Genetic polymorphisms of ARL15 and HLA-DMA are associated with rheumatoid arthritis in Han population from northwest China

目的采用高分辨融解曲线(HRM)技术建立检测类ADP核糖基化因子GTP酶15(ARL15)、主要组织相容性抗原复合体ⅡDMα(HLA-DMA)和核因子κB亚基2(NFKB2)基因单核苷酸多态性(SNP)的方法,并探讨其与中国西北地区汉族人群类风湿性关节炎(RA)易感性的关系。方法针对rs255758、rs1063478、rs397514331和rs397514332四个SNP位点建立PCRHRM检测体系并测序验证。对588例RA患者和200例健康对照标本进行病例对照研究,分析这四个SNP与RA发病风险的关系。结果建立的针对四个SNP位点的PCR-HRM基因分型方法经测序验证可正确分型。rs397514331和rs397514332位点未发现突变基因型。rs255758和rs1063478位点的基因型频率在两组间存在统计学差异,而基因频率无统计学差异。其中rs255758位点的AA基因型在显性模型(AA vs AC/CC)下降低RA发病风险(OR=0.666,95%CI=0.478~0.927,P=0.016)。结论我们建立的PCR-HRM基因分型方法可对rs255758、rs1063478、rs397514331和rs397514332四个SNP位点进行常规化检测。ARL15和HLA-DMA基因多态性与中国西北地区汉族人群RA易感相关。

Objective To establish the methods for detecting single nucleotide polymorphisms（ SNPs） of ADP-ribosylation factor-like GTPase 15（ ARL15）,major histocompatibility complex class Ⅱ-DM alpha（ HLA-DMA） and nuclear factor kappa B subunit 2（ NFKB2） genes using high resolution melting（ HRM） technology,and to explore the association of those SNPs with the susceptibility of rheumatoid arthritis（ RA） in northwestern Han Chinese population. Methods The PCR-HRM detection system for four SNPs（ rs255758,rs1063478,rs397514331 and rs397514332） was established for genotyping,and gene sequencing was performed to validate the genotyping ability of the system. 588 RA cases and 200 controls were enrolled in a case-control study to analyze the associations of ARL15 and HLA-DMA gene polymorphisms with RA risk. Results The direct sequencing validated that the established PCR-HRM detection system could be used for genotyping clinical samples correctly. The mutated genotype of rs397514331 and rs397514332 from NFKB2 gene are not found in this study. The genotype frequencies of rs255758 and rs1063478 had statistical difference between the cases and controls,but no statistical difference in allelic frequencies. Under the dominant model（ AA vs AC/CC）,the AA genotype of rs255758 decreases the RA risk（ OR = 0. 666,95%CI = 0. 478-0. 927,P = 0. 016）. Conclusion The method of PCR-HRM we established can be applied to the routine detection of rs255758,rs1063478,rs397514331 and rs397514332. The ARL15 and HLA-DMA gene polymorphisms are associated with RA risk in Northwestern Han Chinese population.