对源自产志贺样毒素大肠杆菌O157分离株的RAPD-PCR扩增产物进行限制性内切酶分析。

Restriction endonuclease analysis of RAPD-PCR amplicons derived from Shiga-like toxin-producing Escherichia coli O157 isolates.

作者信息

Hopkins Katie L, Hilton Anthony C

机构信息

Department of Biosciences, University of Birmingham, Edgbaston, Birmingham B15 2TT and *Life and Health Sciences, Aston University, Aston Triangle, Birmingham B4 7ET.

出版信息

J Med Microbiol. 2001 Jan;50(1):90-95. doi: 10.1099/0022-1317-50-1-90.

DOI:10.1099/0022-1317-50-1-90

PMID:11192511

Abstract

Shiga-like toxin-producing Escherichia coli O157 isolates were characterised by random amplification of polymorphic DNA by PCR (RAPD-PCR) analysis developed to allow robust epidemiological typing of E. coli. Amplification with primer 1247 or 1290 generated a reproducible profile, but was not capable of distinguishing sufficiently between epidemiologically unrelated strains. Subsequent digestion of the amplicons with selected restriction endonucleases improved the discriminatory ability of this method for strains showing limited differentiation following RAPD-PCR analysis alone. Restriction endonuclease analysis of RAPD-PCR fragments generated from closely related strains has the potential to provide additional discriminatory information without loss of specificity.

摘要

通过聚合酶链反应（PCR）进行随机扩增多态性DNA分析（RAPD-PCR）对产志贺样毒素大肠杆菌O157分离株进行了特征分析，该分析方法旨在对大肠杆菌进行可靠的流行病学分型。用引物1247或1290进行扩增可产生可重复的图谱，但无法充分区分流行病学上无关的菌株。随后用选定的限制性内切酶对扩增产物进行消化，提高了该方法对仅通过RAPD-PCR分析显示有限分化的菌株的鉴别能力。对密切相关菌株产生的RAPD-PCR片段进行限制性内切酶分析，有可能在不丧失特异性的情况下提供额外的鉴别信息。