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通过多重聚合酶链反应鉴定人肠道致病性大肠杆菌菌株

Identification of human enterovirulent Escherichia coli strains by multiplex PCR.

作者信息

Rich C, Alfidja A, Sirot J, Joly B, Forestier C

机构信息

Unité de Bactériologie Moléculaire, Centre Hospitalier Régional Universitaire de Clermont-Ferrand, France.

出版信息

J Clin Lab Anal. 2001;15(2):100-3. doi: 10.1002/jcla.9.

Abstract

Some strains of Escherichia coli are involved in enteric infections in both adults and children. However the classical diagnostic methods can not differentiate pathogenic from nonpathogenic E. coli, because of the lack of phenotypic differences. In this study, we developed multiplex PCR in order to amplify fragments of specific virulence genes of the five main E. coli pathotypes. Fragments of the expected size were obtained using previously or newly designed primers and allowed identification of 10 virulence genes in only 5 reactions. This method was applied to the detection of pathogenic E. coli isolated from 90 patients' stools specimens during an 18-month survey. Patients were suffering from diarrhea or hemolytic uremic syndrome and in 13 cases (14.4%), an enterovirulent E. coli strain was detected. This diagnostic method could therefore represent an important technique in clinical laboratories which lack standard tests for these pathogens.

摘要

一些大肠杆菌菌株会引发成人和儿童的肠道感染。然而,由于缺乏表型差异,传统的诊断方法无法区分致病性大肠杆菌和非致病性大肠杆菌。在本研究中,我们开发了多重聚合酶链反应(multiplex PCR),以扩增五种主要大肠杆菌致病型的特定毒力基因片段。使用先前设计或新设计的引物获得了预期大小的片段,仅通过5次反应就能够鉴定出10个毒力基因。在一项为期18个月的调查中,该方法被应用于检测从90例患者粪便标本中分离出的致病性大肠杆菌。患者患有腹泻或溶血尿毒综合征,在13例(14.4%)中检测到了肠毒性大肠杆菌菌株。因此,这种诊断方法可能成为临床实验室中一项重要的技术,因为这些实验室缺乏针对这些病原体的标准检测方法。

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