应用可视化基因芯片技术检测幽门螺杆菌感染个体化药物治疗相关基因

中国生物工程杂志

2013, Vol. 33

Issue (4): 92-100

技术与方法

应用可视化基因芯片技术检测幽门螺杆菌感染个体化药物治疗相关基因

姚雪¹, 刘琪琦², 赵青¹, 陈苏红²

1. 广州医学院广州 510182;
2. 中国人民解放军军事医学科学院放射与辐射研究所北京 100038

Detection of Individualized Treatment and Test of Helicobacter pylori Infection through Visualized Gene Chip Technology

YAO Xue¹, LIU Qi-qi², ZHAO Qing¹, CHEN Su-hong²

1. Guangzhou Medical College, Guangzhou 510182, China;
2. Beijing Institute of Radiation Medicine, Beijing 100038, China

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摘要： 目的:建立一种能同时检测幽门螺杆菌克拉霉素(23S )、喹诺酮(gyrA)耐药位点和人PPI代谢相关CYP2C19的基因多态性情况的基因芯片检测方法。方法:通过分析筛选人基因组CYP4502C19^*2,CYP4502C19^*3 及幽门螺杆菌克拉霉素和喹诺酮类耐药位点的DNA序列,设计制备了HP感染个体化药物治疗检测基因芯片,并对其特异性、灵敏度、重复性进行了评价。结果:该芯片可以检测出不低于10³ CFU/ml的幽门螺杆菌和不低于2ng/μl 的人基因组DNA。196例临床标本的芯片检测结果与测序结果基本一致。结论:应用可视化基因芯片进行幽门螺杆菌感染个体化药物治疗相关基因检测具有较高的特异性和敏感性,结果准确且易判读,具有较好的应用前景,可以为临床医生提供用药指导。

关键词： 幽门螺杆菌; 耐药; CYP2C19; 多态性; 个体化治疗; 基因芯片; 可视化检测法

Abstract: Objective: To establish a method of visualized gene chip detection that can simultaneously measure the gene polymorphism of CYP2C19 that is related to clarithromycin and quinolones resistance mutation in Helicobacter pylori infection and to the metabolism of proton pump inhibitor. Method: Through analysis the sequence of human CYP2C19^*2, CYP2C19^*3 and the targets related to resistance to clarithromycin and quinolone, a gene chip to test the resistence of Helicobacter Pylori was established. The specificity, sensitivity and repeatability of the gene chip was evaluated in the optimized PCR system, with hybridization reaction and visualized detection. Result: The results of detection of 196 clinical samples using gene chips were consistent with that by sequencing. The detection sensitivity of the chip for Hp was 10³CFU/ml, and that for human genomic DNA was 2ng/μl.Conclusion: This method was quick, highly specific, sensitive, and economic. With its clinical application potential, it is promising in providing medication guidance for Hp elimination in clinical use and individualized treatment.

Key words: Drug resistance of Helicobacter pylori Gene polymorphism of CYP2C19 Individualized treatment Gene chip Visual detection method

收稿日期: 2013-01-07 出版日期: 2013-04-25

ZTFLH:

Q819

基金资助: 国家自然科学基金(81141101);广东省教育部产学研结合项目(2011A090200071);"重大新药创制"科技重大专项(2012ZX09301003-005)资助项目

通讯作者: 赵青, 陈苏红 E-mail: zhaoqing66@yahoo.com;doudoucsh@sina.com

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引用本文:

姚雪, 刘琪琦, 赵青, 陈苏红. 应用可视化基因芯片技术检测幽门螺杆菌感染个体化药物治疗相关基因[J]. 中国生物工程杂志, 2013, 33(4): 92-100.

YAO Xue, LIU Qi-qi, ZHAO Qing, CHEN Su-hong. Detection of Individualized Treatment and Test of Helicobacter pylori Infection through Visualized Gene Chip Technology. China Biotechnology, 2013, 33(4): 92-100.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2013/V33/I4/92

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