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通过聚合酶链反应鉴别志贺毒素和1型维罗细胞毒素基因

Differentiation of Shiga toxin and Vero cytotoxin type 1 genes by polymerase chain reaction.

作者信息

Pollard D R, Johnson W M, Lior H, Tyler S D, Rozee K R

机构信息

National Laboratories for Special and Enteric Pathogens, Laboratory Centre for Disease Control, Ottawa, Ontario, Canada.

出版信息

J Infect Dis. 1990 Nov;162(5):1195-8. doi: 10.1093/infdis/162.5.1195.

Abstract

Two sets of synthetic oligonucleotide primers were used in a polymerase chain reaction technique to distinguish genes for Shiga toxin in Shigella dysenteriae 1 and type 1 Vero cytotoxin (VT1) in Escherichia coli. VT1a and VT1b primers directed at a common 130-base-pair (bp) fragment of the stx and sltI genes detected template nucleic acid in both Shiga toxin-positive S. dysenteriae 1 and VT1-producing E. coli strains. VT1c and VT1d primers, targeting a 140-bp fragment of the promoter region of the sltIA gene, were negative in the polymerase chain reaction with S. dysenteriae 1 nucleic acid and positive with nucleic acids from all strains found to produce VT1 in toxin-specific neutralization tests. Primer specificity was determined in the polymerase chain reaction using nucleic acid extracted from 49 strains of representative enteric pathogens defined in terms of their toxigenicity.

摘要

在聚合酶链反应技术中使用了两组合成寡核苷酸引物,以区分痢疾志贺氏菌1型中的志贺毒素基因和大肠杆菌中的1型维罗细胞毒素(VT1)。针对stx和sltI基因共同的130个碱基对(bp)片段的VT1a和VT1b引物,在产志贺毒素的痢疾志贺氏菌1型和产VT1的大肠杆菌菌株中均检测到模板核酸。靶向sltIA基因启动子区域140 bp片段的VT1c和VT1d引物,在与痢疾志贺氏菌1型核酸的聚合酶链反应中呈阴性,而在毒素特异性中和试验中发现所有产VT1菌株的核酸与之反应时呈阳性。使用从49株根据产毒性定义的代表性肠道病原体中提取核酸,在聚合酶链反应中确定引物特异性。

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