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用于区分来自牛、猪和人类源大肠杆菌的扩增片段长度多态性(AFLP)分析与肠杆菌基因间重复一致序列聚合酶链反应(ERIC-PCR)分析的比较

A comparison of AFLP and ERIC-PCR analyses for discriminating Escherichia coli from cattle, pig and human sources.

作者信息

Leung Kam Tin, Mackereth Rob, Tien Yuan-Ching, Topp Edward

机构信息

Department of Biology, Lakehead University, Thunder Bay, ON, Canada.

出版信息

FEMS Microbiol Ecol. 2004 Jan 1;47(1):111-9. doi: 10.1016/S0168-6496(03)00254-X.

Abstract

Amplified fragment length polymorphism (AFLP) and enterobacterial repetitive intergenic consensus polymerase chain reaction (ERIC-PCR) genomic fingerprinting assays were compared for their ability to differentiate Escherichia coli isolates obtained from various host sources, and with respect to their pathogenicity. One hundred and ten verotoxigenic, enterotoxigenic and non-pathogenic E. coli isolates obtained from cattle, humans and pigs were used in this study. The AFLP assay was shown to be highly effective in predicting both the host source and pathogenicity of the E. coli isolates. A stepwise discriminant function analysis showed that 91.4, 90.6 and 97.7% of the human, bovine and pig isolates were classified into the correct host types, respectively. The analysis also distinguished the non-pathogenic E. coli from the verocytotoxigenic and enterotoxigenic virulence phenotypes at 100, 100 and 90.9% accuracy, respectively. Sixty-two E. coli strains from the collection were subjected to the ERIC-PCR fingerprinting analysis. Using this method, only 28.6, 0 and 75.0% of the human, bovine and pig isolates were classified into the correct host types, respectively. Overall, the AFLP method was able to ascribe host source with a high level of confidence and readily discriminate pathogenic from non-clinical isolates of E. coli.

摘要

对扩增片段长度多态性(AFLP)和肠杆菌重复基因间共识聚合酶链反应(ERIC-PCR)基因组指纹分析方法进行了比较,以评估它们区分从不同宿主来源分离的大肠杆菌菌株的能力及其致病性。本研究使用了从牛、人和猪中分离得到的110株产志贺毒素大肠杆菌、产肠毒素大肠杆菌和非致病性大肠杆菌。结果表明,AFLP分析在预测大肠杆菌菌株的宿主来源和致病性方面都非常有效。逐步判别函数分析显示,分别有91.4%、90.6%和97.7%的人源、牛源和猪源分离株被正确分类到相应的宿主类型中。该分析还分别以100%、100%和90.9%的准确率区分了非致病性大肠杆菌与产志贺毒素大肠杆菌和产肠毒素大肠杆菌的毒力表型。对该菌株库中的62株大肠杆菌进行了ERIC-PCR指纹分析。使用这种方法,分别只有28.6%、0和75.0%的人源、牛源和猪源分离株被正确分类到相应的宿主类型中。总体而言,AFLP方法能够高度可靠地确定宿主来源,并能轻松区分致病性大肠杆菌与非致病性临床分离株。

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