喹诺酮类耐药决定区、外排泵负调控基因对大肠杆菌氟喹诺酮高水平耐药分子机制的影响

›› 2012, Vol. 39 ›› Issue (10): 11-16.

喹诺酮类耐药决定区、外排泵负调控基因对大肠杆菌氟喹诺酮高水平耐药分子机制的影响

任艳娜, 甄盼盼, 李健, 郭玉芳, 王丽华, 任君玉, 张文慧, 蒋红霞

华南农业大学兽医学院,广东省兽药研制与安全评价重点实验室,广东广州 510642

收稿日期:2012-02-07 出版日期:2012-10-20 发布日期:2012-10-19
通讯作者: 蒋红霞(1968－),女,副教授,研究方向:细菌耐药性、兽药安全评价。Tel: 020-85283934;Fax: 020-85284896;E-mail:hxjiang@scau.edu.cn E-mail:hxjiang@scau.edu.cn
作者简介:任艳娜(1985－),女,河南人,硕士生,研究方向:动物细菌耐药机制。
基金资助:
国家自然科学基金—广东省自然科学基金联合基金重点项目(U1031004,U0631006)。

Genetic Characterization of Highly Fluoroquinolone Resistant Clinical Escherichia coli Strains:Role of QRDR,acrR,marR and soxR

REN Yan-na, ZHEN Pan-pan, LI Jian, GUO Yu-fang, WANG Li-hua, REN Jun-yu, ZHANG Wen-hui, JIANG Hong-xia

Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation, College of Veterinary Medicine, South China Agricultural University, Guangzhou 510642,China

Received:2012-02-07 Online:2012-10-20 Published:2012-10-19

摘要/Abstract

摘要： 为了探讨耐喹诺酮类决定区(QRDR)、外排泵负调控基因(acrR、marR和soxR)突变对临床分离株氟喹诺酮(FQs)高水平耐药的影响,本研究对临床分离的18株FQs耐药大肠杆菌(E.coli),采用PCR方法检测QRDR、acrR、marR和soxR的突变情况;通过RT-PCR的方法检测外排泵及膜孔蛋白相关基因的表达水平。结果显示,QRDR的突变主要集中在常规突变GyrA(Ser83Leu 和 Asp87Asn)和ParC(Ser80Ile),同时也检测出稀有突变ParC Glu84Gly、Glu84Lys、Glu84Val和Glu84Ala,ParE Ser458Ala等。ED28在acrR基因存在777 bp插入序列;12株菌(包括ATCC25922)在MarR存在Gly103Ser和Tyr137His双突变,其中EP26和EG42存在插入片段;ED40在SoxR存在Thr38Ser、Gly74Arg氨基酸替换。在多突变药菌株中,AcrAB的表达水平明显升高,OmpC和OmpF表达量降低、甚至缺失。

关键词: QRDR; acrR; marR; soxR; 耐药; 氟喹诺酮

Abstract: The objective of this study was to detect the role of QRDR, acrR, marR and soxR in genetic characterization of highly fluoroquinolone-resistant clinical Escherichia coli strains. PCR method was used to analyse the mutations of QRDR, acrR, marR and soxR. RT-PCR method was used to detect the expression level of efflux pump and porin related genes. Mutations of QRDR mainly fell into GyrA(Ser83Leu and Asp87Asn) and ParC(Ser80Ile), and other mutations had also detected, such as ParC Glu84Gly, Glu84Lys, Glu84Val and Glu84Ala,ParE Ser458Ala,et al. There was an insertion of 777 bp fragment within acrR in ED28. 12 of 18 stains including ATCC25922 had double mutations (Gly103Ser and Tyr137His) in MarR and two (EP26 and EG42) of them had an insertion. ED40 had amino acid substitution (Thr38Ser and Gly74Arg) in SoxR. In multiple mutations strains, there were markedly increased expression of AcrAB and decrease expression of porin or porin loss.

Key words: QRDR; acrR; marR; soxR; resistance; fluoroquinolone

中图分类号:

S852.61⁺4

任艳娜, 甄盼盼, 李健, 郭玉芳, 王丽华, 任君玉, 张文慧, 蒋红霞. 喹诺酮类耐药决定区、外排泵负调控基因对大肠杆菌氟喹诺酮高水平耐药分子机制的影响[J]. , 2012, 39(10): 11-16.

REN Yan-na, ZHEN Pan-pan, LI Jian, GUO Yu-fang, WANG Li-hua, REN Jun-yu, ZHANG Wen-hui, JIANG Hong-xia. Genetic Characterization of Highly Fluoroquinolone Resistant Clinical Escherichia coli Strains:Role of QRDR,acrR,marR and soxR[J]. China Animal Husbandry & Veterinary Medicine, 2012, 39(10): 11-16.

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