Tetra-primer ARMS PCR检测丙型肝炎患者IL-28B rs12979860基因多态性

Tetra-primer ARMS PCR for detecting polymorphisms of IL-28B gene rs12979860 in patients with hepatitis C

目的评价四引物扩增受阻突变体系聚合酶链反应(Tetra-primer ARMS PCR)检测IL-28Brs12979860基因多态性在丙型肝炎患者抗病毒治疗中的应用。方法选取2015年5月至2016年7月就诊的慢性丙型肝炎患者275例,提取外周血DNA,建立Tetra-primer ARMS PCR检测体系,与Sanger测序法对比验证,检测该地区丙型肝炎患者IL-28Brs12979860基因多态性分布情况。结果 Tetra-primer ARMS PCR检测体系成功建立,与Sanger测序法的符合率100%,rs12979860基因的C/C型、C/T型和T/T型的分布频率分别为86.5%、12.0%、1.5%。在C/C型组与C/T或T/T型组中,年龄分布、性别分布、HCV基因1b型比例差异均无统计学意义(P>0.05);患者肝硬化比例和持续病毒学应答比例比较差异有统计学意义(P<0.05),在C/T或T/T组中肝硬化比例较高,在C/C组中持续病毒学应答比例较高。结论该研究建立的Tetra-primer ARMS PCR技术是一种操作简单、成本低廉、快速有效的基因多态性检测方法,对于丙型肝炎患者IL-28Brs12979860基因多态性基因分型具有很好的临床应用价值。

Objective To evaluate the application of tetra-primer amplification refractory mutation system PCR（Tetra primer ARMS PCR） fro detecting IL-28B rs12979860 gene polymorphism in the patients with hepatitis C. Methods Two hundreds and seventy-five patients with chronic hepatitis C in our hospital from May 2015 to July 2016 were selected and their peripheral blood DNA was extracted. The detection system of Tetra-primer ARMS PCR was established,which was compared with the Sanger se- quencing. Then this technique was used to detect IL-28B rs12979860 gene polymorphism in the patients with hepatitis C in this are- a. Results The detection system of Tetra-primer ARMS PCR was successfully established with a consistent rate of 100% with the Sanger sequencing. The distribution frequencies of C/C,C/T and T/T of IL-28B rs12979860 gene were 86.5 %, 12.0 % and 1.5 respectively. The age distribution, gender distribution and HCV genotype 1b ratio had no statistically significant difference（P〉0.05）, however,the rate of patients with liver cirrhosis has a statistical difference between the genotype C/C group with the geno- type T/T group or genotype C/T group（P〈0.05） ;while the proportion of liver cirrhosis and sustained virological response had the statistical difference（P〈0.05） ,the proportion of liver cirrhosis in the genotype C/T group or T/T group was higher,and the pro- portion of sustained virological response in the genotype C/C group was higher. Conclusion The Tetra-primer ARMS PCR tech- nique established by this research is simple to operate,low cost, fast and effective detection method of gene polymorphism, which has a better clinical application value in the genotyping of IL-28B rs12979860 gene polymorphism in the patients with hepatitis C.