文章摘要

福氏志贺氏菌QRDR基因gyrA,parC及毒力基因ipaH的多重PCR检测

作者: 1王 倩, 1徐 新明, 1胡 蕊, 1谢 常宝, 1王 尹, 2王 鑫, 1顾 兵, 1陈 莹
1 徐州医科大学医学技术学院,江苏 徐州 221004
2 南京巨鲨显示科技有限公司,江苏 南京 210036
通讯: 陈 莹 Email: cyylotus@163.com
DOI: 10.3978/j.issn.2095-6959.2019.01.004
基金: 国家自然科学基金(81702103);江苏省自然科学基金(BK20170252);江苏省高校自然科学研究项目(16KJD320005);江苏省高校大学生创新创业训练计划(201610313019Z);徐州市科技计划项目(KC16SY15)。

摘要

目的:建立快速检测福氏志贺菌致病性、耐药性的多重PCR方法。方法:根据福氏志贺菌耐药性基因gyrA,parC及毒力致病性基因ipaH,分别设计了3对引物,其PCR扩增产物分别为648,248和423 bp,并进行单基因PCR和单管多重PCR扩增特异性和Tm值的优化。结果:该方法可同时检测福氏志贺菌gyrA,parC及ipaH三种标志性靶基因,其最优退火温度为57.5 ℃。结论:本方法操作简单,检测周期短,能够实现对福氏志贺菌样本的喹诺酮耐药决定区基因、毒力基因的并行快速检测。
关键词: 多重PCR;福氏志贺菌;耐药基因;致病基因

Multiplex PCR detection of QRDR genes gyrA, parC and virulence gene ipaH of Shigella flexneri

Authors: 1WANG Qian, 1XU Xinming, 1HU Rui, 1XIE Changbao, 1WANG Yin, 2WANG Xin, 1GU Bing, 1CHEN Ying
1 School of Medical Technology, Xuzhou Medical University, Xuzhou 221004, China
2 Nanjing Jusha Display Technology Co., Ltd, Nanjing 210036, China

CorrespondingAuthor:CHEN Ying Email: cyylotus@163.com

Foundation: This work was supported by the National Natural Science Foundation (81702103), Jiangsu Provincial Natural Science Foundation (BK20170252)

Abstract

Objective: To establish a multiplex PCR method for rapid detection of pathogenicity and drug resistance of Shigella flexneri. Methods: Three pairs of primers were designed according to the gyrA, parC and the virulence pathogenicity gene ipaH of Shigella flexneri, and their PCR amplification products were 648, 248 and 423 bp, respectively, and single gene PCR and single tube were performed. Multiplex PCR amplification specificity and Tm value optimization were optimized. Results: The method can parallel detect three target genes of gyrA, parC and ipaH of Shigella flexneri, and the optimal annealing temperature is 57.5 ℃. Conclusion: This method is simple and has a short detection cycle. It can rapidly parallel detect QRDR genes and virulence gene of Shigella flexneri.
Keywords: multiplex PCR; Shigella flexneri; drug resistance gene; pathogenicity gene