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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。
Abstract BACKGROUND: Polymorphisms of cytochrome P450 isoenzyme 2C19 (CYP2C19) and interleukin-1β (IL-1β) are important influencing factors for the eradication rate of Helicobacter pylori (Hp) infection and their etiological factors The development of treatment plan is instructive. OBJECTIVE: To identify genotypes of single nucleotide polymorphisms (SNPs) of CYP2C19 and IL-1β using high resolution melting curve (HRM) technique. Methods: Peripheral blood samples from 200 healthy volunteers were collected and genomic DNA was extracted. The optimal Mg2 + concentrations of CYP2C19 * 2, * 3 and IL-1β-31 / -511 primers were determined by HRM and the genotypes of the corresponding SNPs were detected. The test results are verified by gene sequencing. Results: The optimal concentrations of Mg2 + for CYP2C19 * 2, * 3 and IL-1β-31 / -511 were 3.0, 3.0, 2.5 and 2.5 mmol / L, respectively. HRM technology can accurately identify CYP2C19 * 2, * 3 and IL-1β-31 / -511 genotypes and has good consistency with the gene sequencing results (κ = 0.985 and 0.968). Conclusion: HRM can accurately identify CYP2C19 and IL-1β gene polymorphisms and can be used to guide the choice of drugs for Hp eradication therapy.