Pollard D R, Johnson W M, Lior H, Tyler S D, Rozee K R
National Laboratories for Special and Enteric Pathogens, Laboratory Centre for Disease Control, Ottawa, Ontario, Canada.
J Infect Dis. 1990 Nov;162(5):1195-8. doi: 10.1093/infdis/162.5.1195.
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.
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。
