利用同一份胃黏膜标本进行幽门螺杆菌分离培养及 CYP2 C19基因多态性检测的研究

摘要

目的：分析北京和山东地区胃病患者CYP2C19代谢类型分布，以及探索用一份胃黏膜标本同时进行幽门螺杆菌（ Helicobacter pylori，HP）分离培养及CYP2C19基因多态性检测的可行性。方法2013年3月至2014年4月收集北京和山东地区快速尿素酶试验（RUT）阳性患者胃黏膜活检标本共1649份，利用同一份胃黏膜活检标本研磨后进行HP菌株分离培养和提取胃黏膜标本基因组DNA用于患者 CYP2C19基因多态性检测。 CYP2C19基因多态性检测中，利用 CYP2C19倡2、CYP2C19倡3特异性引物进行PCR扩增，将PCR产物进行琼脂糖凝胶电泳检测，分析成功率并进行DNA测序，并分析北京和山东地区CYP2C19代谢类型分布情况。结果1649份胃黏膜标本成功分离HP菌株1415株，分离率为85．81％。 CYP2C19特异PCR扩增产物经琼脂糖凝胶电泳条带清晰特异，成功率为100％且符合测序要求。统计分析结果显示北京地区和山东地区胃病患者CYP2C19代谢类型间差异无统计学意义（P＞0．05），性别对于北京和山东地区CYP2C19基因多态性分布无显著性影响。结论利用同一份胃黏膜标本进行HP菌株分离培养及CYP2C19基因多态性检测具有可行性。北京地区与山东地区胃病患者 CYP2 C19代谢类型之间差异无统计学意义且性别对于CYP2C19基因多态性分布无显著性影响。%Objective To analyze the CYP2C19 genetic polymorphisms in patients suffering from gastric diseases in Beijing and Shandong, and to evaluate the possibility of using the same gastric biopsy samples for the isolation of Helicobacter pylori ( H.pylori) strains.Methods A total of 1649 gastric biopsy samples positive for rapid urease test ( RUT) were collected from patients with gastric diseases in Beijing and Shandong.Each sample was grinded and equally divided into two parts for the isolation of H.pylori strains and the analysis of CYP2C19 genetic polymorphisms, respectively.H.pylori strains were isolated from the samples and identified with catalase and urease.CYP2C19*2 and CYP2C19*3 specific primers were used for the PCR amplification of DNA extracts.The PCR products were analyzed by electrophoresis and nucleo-tide sequencing.The metabolic types of CYP2C19 were analyzed based on the results of nucleotide sequen-cing.Results Altogether 1415 (85.81%) H.pylori strains were isolated successfully from the 1649 gastric biopsy samples.The products of CYP2C19 specific PCR for all DNA extracts showed clear bands in electro-phoresis and were further confirmed with sequencing analysis.Statistical analysis revealed that there were no statistical differences with the percentages of CYP2C19 metabolic types between Beijing and Shandong, nor between males and females (P>0.05).Conclusion It showed the possibility of using the same gastric bi-opsy samples for the isolation of H.pylori strains and the analysis of CYP2C19 genetic polymorphisms.No significant differences with the percentages of CYP2C19 metabolic types were found between patients from Beijing and Shandong.