【摘要】：【目的】研究廢水中產(chǎn)超廣譜β-內(nèi)酰胺酶大腸桿菌中可移動質(zhì)粒在耐藥基因水平傳播機制中的作用�！痉椒ā繉ξ鬯畯S分離所得的50株產(chǎn)ESBLs大腸桿菌進行接合試驗,并對所得的接合子采用紙片擴散法測定其對15種常見藥物的耐藥表型,針對質(zhì)粒介導的產(chǎn)ESBLs菌株的耐藥基因設(shè)計7對特異性引物對接合子進行PCR擴增。【結(jié)果】研究結(jié)果顯示,80份水樣分離得50株產(chǎn)ESBLs大腸桿菌,共接合成功35株細菌,接合成功率高達70%。接合子與供體菌相比,均發(fā)生耐藥譜型的改變,且存在丟失一種或幾種藥物耐藥性且產(chǎn)生另一種或幾種藥物耐藥性的現(xiàn)象。PCR擴增結(jié)果顯示,接合子與供體菌相比,耐藥基因型有所減少或不變,bla_(TEM)、bla_(CTX-M)基因全部接合成功,bla_(SHV)基因僅1株未接合成功,耐氟喹諾酮類基因未發(fā)生轉(zhuǎn)移。【結(jié)論】本研究表明,不同的耐藥基因可能位于不同的可移動質(zhì)粒上,可移動質(zhì)粒在大腸桿菌耐藥性水平傳播的過程中起到了十分重要的作用。
[Abstract]:[objective] to study the role of moveable plasmid in extended-spectrum 尾-lactamase-producing Escherichia coli in the horizontal transmission mechanism of drug-resistant genes in wastewater. [methods] 50 strains of ESBLs-producing Escherichia coli isolated from sewage plants were used for conjugation test. The resistance phenotype to 15 common drugs was determined by disk diffusion method. Seven pairs of specific primers were designed to amplify the resistant gene of plasmid-mediated ESBLs-producing strain by PCR. [results] the results showed that the resistant phenotype to 15 kinds of common drugs was detected by PCR. [results] the results of this study showed that the resistant phenotype to 15 common drugs was amplified by PCR with 7 pairs of primers. Fifty strains of ESBLs-producing Escherichia coli were isolated from 80 water samples. 35 strains of E. coli were successfully conjugated, and the successful rate of conjugation was up to 70%. Compared with donor bacteria, the heterozygotes showed changes in drug resistance patterns, and the loss of one or more drug resistance and the production of another or more drug resistance. PCR results showed that compared with donor bacteria, the heterozygote had the same drug resistance as donor bacteria, and the results of PCR amplification showed that compared with donor bacteria, the heterozygote had one or more drug resistance. The genotype of drug resistance was decreased or unchanged, the bla_ (TEM), bla_ (CTX-M) gene was successfully joined, the bla_ (SHV) gene was not successfully joined, and the fluoroquinolone resistant gene was not transferred. [conclusion] the present study indicates that the gene of fluoroquinolone is not transferred. Different drug resistance genes may be located on different moveable plasmids, which play a very important role in the level of drug resistance transmission in E. coli.
【作者單位】： 山東農(nóng)業(yè)大學動物科技學院;山東省農(nóng)業(yè)科學院;泰山醫(yī)學院基礎(chǔ)醫(yī)學部;
【基金】：基金項目:山東省農(nóng)業(yè)產(chǎn)業(yè)技術(shù)創(chuàng)新體系家禽創(chuàng)新團隊(SDA/T-11-09)~~
【分類號】：R378
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本文編號：2456991