大腸桿菌O157∶H7對氟苯尼考耐藥機制的初步研究
發(fā)布時間:2018-04-03 14:26
本文選題:大腸桿菌O∶H 切入點:氟苯尼考 出處:《中國人獸共患病學(xué)報》2016年03期
【摘要】:目的研究大腸桿菌O157∶H7(E.coli O157∶H7)對氟苯尼考耐藥性產(chǎn)生的機制。方法本研究采用亞抑菌濃度體外耐藥誘導(dǎo)的方法將E.coli O157∶H7誘導(dǎo)成為氟苯尼考高度耐藥菌株,采用無抗生素壓力下連續(xù)傳代培養(yǎng)或SDS方法將耐藥菌株的氟苯尼考耐藥性消除,使用PCR方法檢測氟苯尼考耐藥基因floR,通過外排泵抑制劑與氟苯尼考聯(lián)合使用,檢測E.coli O157∶H7是否存在對氟苯尼考的主動外排作用。結(jié)果經(jīng)過36代的耐藥誘導(dǎo),敏感菌株對氟苯尼考產(chǎn)生了高度耐藥(MIC為256μg/mL)。質(zhì)粒檢測結(jié)果顯示,耐藥誘導(dǎo)菌株出現(xiàn)2個質(zhì)粒,大小分別約為3 500bp和1 200bp,floR基因位于質(zhì)粒DNA中。經(jīng)連續(xù)傳代培養(yǎng)或SDS法進行耐藥消除,耐藥誘導(dǎo)菌株對氟苯尼考的敏感性提高,但其質(zhì)粒和floR基因均未丟失。結(jié)論 E.coli O157∶H7氟苯尼考敏感菌和耐藥菌均存在對氟苯尼考的質(zhì)子驅(qū)動型外排泵和ATP水解依賴型外排泵主動外排作用。
[Abstract]:Objective to study the mechanism of drug resistance of Escherichia coli O157:H7(E.coli O 157: h 7 to florfenicol.Methods in this study, E.coli O157:H7 was induced into florfenicol high resistance strain by subinhibitory concentration in vitro. The drug resistance of florfenicol resistant strains was eliminated by continuous passage culture without antibiotic pressure or by SDS method.Florfenicol resistance gene flor was detected by PCR method. The efflux pump inhibitor combined with florfenicol was used to detect whether E.coli O157:H7 had active efflux effect on florfenicol.Results after 36 generations of drug resistance induction, the sensitive strain produced high resistance to florfenicol with a MIC of 256 渭 g / mL 路L ~ (-1).The results of plasmid detection showed that two plasmids, about 3 500bp and 1 200 BP flor gene, were found in plasmid DNA.The sensitivity of the drug-resistance induced strains to florfenicol was improved by continuous passage culture or SDS method, but neither the plasmid nor the floR gene was lost.Conclusion both E.coli O157:H7 florfenicol sensitive bacteria and drug resistant bacteria have proton driven efflux pump and ATP hydrolytic dependent efflux pump active efflux.
【作者單位】: 廣西大學(xué)動物科學(xué)技術(shù)學(xué)院;廣西獸醫(yī)研究所;廣西畜禽疫苗新技術(shù)重點實驗室;
【基金】:廣西科技重大專項(桂科重14121003-4-1) 廣西自然科學(xué)基金(桂科自2013GXNSFAA019093) 廣西水產(chǎn)畜牧獸醫(yī)局科研專項(桂漁牧科201528005)~~
【分類號】:R378
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本文編號:1705587
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