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用于检测和鉴定产肠毒素大肠杆菌的基因分型检测方法的开发与评估

Development and evaluation of genotypic assays for the detection and characterization of enterotoxigenic Escherichia coli.

作者信息

Steinsland Hans, Valentiner-Branth Palle, Grewal Harleen M S, Gaastra Wim, Mølbak K Kåre, Sommerfelt Halvor

机构信息

Centre for International Health, University of Bergen, Norway.

出版信息

Diagn Microbiol Infect Dis. 2003 Feb;45(2):97-105. doi: 10.1016/s0732-8893(02)00504-7.

Abstract

We developed and evaluated a method to genotypically identify enterotoxigenic Escherichia coli (ETEC) and to characterize these organisms with respect to 18 of 21 known colonization factors (CFs). The method, which is based on polynucleotide DNA-DNA colony hybridization, includes a pooled toxin probe assay to identify ETEC, and individual probe assays to detect the enterotoxins STp, STh, and LT, and the CFs CFA/I, CS1-CS8, CS12-CS15, CS17-CS19, CS21, and CS22. We evaluated the pooled toxin probe assay during a cohort study of childhood diarrhea, and the individual probe assays against 33 reference strains and 92 clinical ETEC isolates. There was close to a complete agreement between the pooled toxin probe assay and the individual toxin probe assays, and between the individual CF probe assays and the corresponding phenotypic assays.

摘要

我们开发并评估了一种方法,用于从基因分型上鉴定产肠毒素大肠杆菌(ETEC),并针对21种已知定植因子(CFs)中的18种对这些微生物进行特征描述。该方法基于多核苷酸DNA-DNA菌落杂交,包括用于鉴定ETEC的混合毒素探针检测,以及用于检测肠毒素STp、STh和LT,以及CFs CFA/I、CS1-CS8、CS12-CS15、CS17-CS19、CS21和CS22的单个探针检测。我们在一项儿童腹泻队列研究中评估了混合毒素探针检测,并针对33株参考菌株和92株临床ETEC分离株评估了单个探针检测。混合毒素探针检测与单个毒素探针检测之间,以及单个CF探针检测与相应的表型检测之间几乎完全一致。

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