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通过随机扩增多态性DNA对禽源大肠杆菌分离株进行分子分型

Molecular typing of avian Escherichia coli isolates by random amplification of polymorphic DNA.

作者信息

Maurer J J, Lee M D, Lobsinger C, Brown T, Maier M, Thayer S G

机构信息

Department of Avian Medicine, University of Georgia, Athens, USA.

出版信息

Avian Dis. 1998 Jul-Sep;42(3):431-51.

PMID:9777144
Abstract

Escherichia coli is a common inhabitant of the gastrointestinal tract of most animals. Like most pathogenic E. coli, avian isolates cannot be distinguished biochemically from the normal commensals inhabiting the gastrointestinal tract of birds. Using a molecular approach, we were able to identify genetic differences among avian E. coli isolates by restriction fragment length polymorphism (RFLP) and random amplification of polymorphic DNA (RAPD) by the polymerase chain reaction (PCR). Several different RFLPs were observed among avian E. coli isolates using DNA probes for 16S ribosomal RNA genes (rrn) and insertion sequence elements (IS2). We were also able to observe differences in DNA banding patterns generated by RAPD analysis. Similarities and differences among avian E. coli were discernible using RFLPs and RAPD analysis, whereas conventional bacteriological methods failed to differentiate these isolates. Based on RAPD patterns, avian E. coli appear to be genetically diverse. Of 16 different RAPD types (RT) encountered, 84% of E. coli fell into seven major RTs. One RT was present in clinical isolates but absent from the commensals isolated in this study. Many of these different E. coli RTs were not geographically restricted to northern Georgia but were also observed in other southern states in the United States. Resistance to various antibiotics was randomly associated with different E. coli RTs. Sarafloxacin resistance was present among different E. coli RTs, suggesting that antibiotic usage is not selecting for a clonal population in avian E. coli. RAPD provides a rapid and powerful tool to study the epidemiology of avian E. coli.

摘要

大肠杆菌是大多数动物胃肠道的常见寄居菌。与大多数致病性大肠杆菌一样,禽源分离株在生化特性上无法与栖息于鸟类胃肠道的正常共生菌区分开来。我们采用分子方法,通过聚合酶链反应(PCR)的限制性片段长度多态性(RFLP)和随机扩增多态性DNA(RAPD)技术,能够鉴定禽源大肠杆菌分离株之间的遗传差异。使用针对16S核糖体RNA基因(rrn)和插入序列元件(IS2)的DNA探针,在禽源大肠杆菌分离株中观察到了几种不同的RFLP。我们还能够观察到RAPD分析产生的DNA条带模式差异。使用RFLP和RAPD分析可以辨别禽源大肠杆菌之间的异同,而传统的细菌学方法无法区分这些分离株。基于RAPD模式,禽源大肠杆菌似乎具有遗传多样性。在遇到的16种不同RAPD类型(RT)中,84%的大肠杆菌属于7种主要RT。其中一种RT存在于临床分离株中,但在本研究分离的共生菌中不存在。许多这些不同的大肠杆菌RT在地理上并不局限于佐治亚州北部,在美国其他南部州也有观察到。对各种抗生素的耐药性与不同的大肠杆菌RT随机相关。不同的大肠杆菌RT中均存在对沙拉沙星的耐药性,这表明抗生素的使用并未在禽源大肠杆菌中选择出克隆群体。RAPD为研究禽源大肠杆菌的流行病学提供了一种快速而强大的工具。

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