新疆多源喹诺酮类耐药大肠杆菌耐药基因检测及分析

doi:10.3864/j.issn.0578-1752.2014.20.018

摘要/Abstract

摘要： 【目的】了解新疆不同动物源喹诺酮类耐药大肠杆菌携带质粒介导的喹诺酮耐药因子（plasmid mediated quinolone resistance，PMQR）的流行现状，及其与主要的β-内酰胺酶基因和 16S rRNA 甲基化酶基因的共存情况。【方法】通过 PCR 方法对猪源（79 株）、牛源（8株）和羊源（96株）喹诺酮类（环丙沙星，诺氟沙星和恩诺沙星）耐药大肠杆菌进行 PMQR（qnrA, qnrB, qnrC, qnrD, qnrS, qepA, oqxA, oqxB, aac(6’)-Ib-cr）、β-内酰胺酶基因（bla_TEM, bla_CTX-M, bla_SHV, bla_KPC, bla_CMY-2, bla_LAP-1）及16S rRNA（armA, rmtB）等基因检测，对目的条带进行DNA测序，确定阳性菌株，对检出携带 β-内酰胺酶基因和16S rRNA 甲基化酶基因的大肠杆菌进行β-内酰胺类及氨基糖苷类药敏试验，分析其携带的基因型与耐药表型之间的关系。【结果】猪源大肠杆菌中检出的PMQR因子为qnrS（5/79，6.33%），oqxA（35/79，44.30%），oqxB（40/79，50.60%）和aac(6’)-Ib-cr（4/79，5.06%），检出的β-内酰胺酶基因为bla_TEM（79/79，100%），检出的16S rRNA 基因为rmtB（3/79，3.80%）；牛源大肠杆菌中检出的PMQR因子为qnrS（1/8，12.50%），oqxA（1/8，12.50%），oqxB（1/8，12.50%）和aac(6’)-Ib-cr（1/8，12.50%）和qepA（1/8，12.50%），检出的β-内酰胺酶基因为bla_TEM（8/8，100%）和bla_SHV（1/8，12.50%）；羊源大肠杆菌中检出的PMQR因子为qnrS（6/96，6.25%），oqxA（32/96，33.3%），oqxB（37/96，38.5%）和aac(6’)-Ib-cr（22/96，22.91%），检出的β-内酰胺酶基因为bla_TEM（96/96，100%）和bla_CTX-M（2/96，2.08%），检出的16S rRNA 基因为rmtB（2/96，2.08%）；未检出qnrA，qnrB，qnrC，qnrD，bla_KPC，bla_CMY-2和bla_LAP-1被检基因。不同动物源大肠杆菌中存在基因共存现象，携带β-内酰胺酶基因和16S rRNA 甲基化酶基因的喹诺酮类耐药大肠杆菌对β-内酰胺类和氨基糖苷类药物耐药呈一定的相关性。【结论】不同动物源喹诺酮类耐药大肠杆菌中存在PMQR因子，可与主要的 β-内酰胺酶和/或16S rRNA 甲基化酶基因共存。此外，首次在羊源大肠杆菌中检出PMQR 因子、β-内酰胺酶及16S rRNA甲基化酶基因。

关键词: 喹诺酮类耐药大肠杆菌, 不同动物源, PMQR因子, 检测

Abstract: 【Objective】The objective of this study is to investigate the prevalence and characteristics of plasmid mediated quinolone resistance (PMQR) determinents in quinolone-resistant E. coli from different animal origins in Xinjiang, and their coexistence with the major β-lactamases and 16s rRNA methylation enzyme genes. 【Method】 Polymerase chain reaction (PCR) was used to detect PMQR (qnrA, qnrB, qnrC, qnrD, qnrS, qepA, oqxA, oqxB, aac(6’)-Ib-cr), b-lactamase gene (bla_TEM, bla_CTX-M, bla_SHV, bla_KPC, bla_CMY-2, bla_LAP-1) and 16S rRNA (armA, rmtB) genes in 79 strains from pigs, 8 strains from cattle and 96 strains from sheep quinolone-resistant (ciprofloxacin, norfloxacin, enrofloxacin) E. coli. The positive strains were performed by using DNA sequencing to determine the purpose of the belt.Minimal inhibitory concentrations (MICs) of the related antibiotics to these isolates carrying the β-lactamase and 16S rRNA methylase genes,were tested and the relationship between the genotype and resistant phenotype was analyzed.【Result】The results showed that the qnrS (6.33%, 5/79), aac(6')-Ib-cr (5.06%, 4/79), oqxA (44.3%, 35/79), oqxB (50.6%, 40/79) were main PMQR determinants, bla_TEM (100%, 79/79) was main β-lactamase genes and rmtB (3.80%, 3/79) was main16S rRNA methylase genes in E. coli from pigs; the qnrS (12.50%, 1/8), oqxA (12.5%, 1/8), oqxB (12.5%,1/8), aac(6')-Ib-cr (12.50%, 1/8) and qepA (12.50%, 1/8) were main PMQR determinants, the bla_TEM (100%, 8/8) and bla_SHV (12.50%, 1/8) were main β-lactamase genes in E. coli from cattle; the qnrS (6.25%, 6/96), oqxA (33.3%, 32/96) oqxB (38.5%, 37/96) and aac(6')-Ib-cr (22.91%, 22/96) were main PMQR determinants, the bla_TEM (100%, 96/96) and bla_CTX-M (2.08%, 2/96) were main β-lactamase genes and the rmtB (2.08%, 2/96) was main16S rRNA methylase genes in E. coli from sheep. The qnrA, qnrB, qnrC, qnrD, bla_KPC, bla_CMY-2 and bla_LAP-1 genes were not detected in any of the isolates. The co-harboring of resistant genes was common among these E.coli from different animals, and the resistant phenotype of E.coli from different animal sources carrying β-lactam enzyme and 16S rRNA methylase genes has some correlation.【Conclusion】The PMQR determinants were existed in E. coli from different animals in Xinjiang, and they coexist with the main b-lactamase and/or 16s rRNA methylation enzyme genes. In addition, the PMQR determinants and β-lactamase genes were firstly detected in the E. coli from sheep.

Key words: quinolone-resistant E. coli, different animal origin, PMQR determinants, detection

南海辰，底丽娜，夏利宁. 新疆多源喹诺酮类耐药大肠杆菌耐药基因检测及分析[J]. 中国农业科学, 2014, 47(20): 4096-4108.

NAN Hai-chen, DI Li-na, XIA Li-ning. Detection and Analysis of Resistance Genes in Quinolone-Resistant Escherichia coli Isolates from Different Livestocks in Xinjiang[J]. Scientia Agricultura Sinica, 2014, 47(20): 4096-4108.

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