基于高分辨率熔解曲线技术的CYP2C19、IL-1β基因多态性检测

Identification of CYP2C19 and IL-1β Polymorphisms Based on High-resolution Melting Curve Analysis

背景:细胞色素P450同工酶2C19(CYP2C19)和白细胞介素-1β(IL-1β)基因多态性是幽门螺杆菌(Hp)感染根除率的重要影响因素,明确其基因多态性对根除治疗方案的制定具有指导意义。目的:采用高分辨率熔解曲线(HRM)技术鉴别CYP2C19、IL-1β基因单核苷酸多态性(SNPs)位点基因型。方法:采集200名健康体检者外周血,提取基因组DNA,以HRM技术确定CYP2C19*2、*3和IL-1β-31/-511引物的最佳Mg2+浓度,并检测相应SNPs位点基因型,检测结果以基因测序进行验证。结果:CYP2C19*2、*3和IL-1β-31/-511引物的最佳Mg2+浓度分别为3.0、3.0、2.5、2.5 mmol/L。HRM技术能精确鉴别CYP2C19*2、*3和IL-1β-31/-511基因型,与基因测序结果有良好的一致性(κ=0.985和0.968)。结论:HRM技术能精确鉴别CYP2C19、IL-1β基因多态性,可用于指导Hp根除治疗方案的药物选择。

Background： The eradication rate of Helicobacter pylori（ Hp） infection is strongly influenced by cytochrome P4502C19（ CYP2C19） and interleukin-1β（ IL-1β） polymorphisms,identification of these gene polymorphisms may provide a guidance for the selection of eradication regimen. Aims： To identify the genotypes of single nucleotide polymorphisms（ SNPs） in CYP2C19 and IL-1β genes by using high-resolution melting curve（ HRM） analysis. Methods： Peripheral blood was obtained from 200 healthy subjects and the genomic DNA was extracted. The optimal concentrations of Mg^2 ＋for primers of CYP2C19＊ 2,＊ 3 and IL-1β-31 /-511 were determined by HRM analysis. Then,the genotypes of SNPs in CYP2C19 and IL-1β were identified by HRM analysis and verified by gene sequencing. Results： The optimal concentrations of Mg^2 ＋for primers of CYP2C19 ＊ 2,＊ 3 and IL-1β-31 /-511 were 3. 0,3. 0,2. 5 and 2. 5 mmol / L,respectively. CYP2C19 ＊ 2,＊ 3 and IL-1β-31 /-511 genotypes could be accurately identified by HRM analysis,which had excellent consistency with the results of gene sequencing（ κ = 0. 985 and 0. 968,respectively）. Conclusions： CYP2C19 and IL-1β polymorphisms can be accurately identified by HRM analysis,which might be used as a reference for the selection of Hp eradication regimen.