Woodward M J, Carroll P J, Wray C
Department of Bacteriology, New Haw, Weybridge, UK.
Vet Microbiol. 1992 Jun 1;31(2-3):251-61. doi: 10.1016/0378-1135(92)90083-6.
Oligonucleotide primers were designed for the specific polymerase chain reaction (PCR) amplification of the enterotoxins STIa and LTI and of the verocytotoxins VT1 and VT2. All of 184 E. coli isolates from cases of diarrhoea from pigs, cattle and sheep gave identical toxin profiles by PCR and gene probe. Differentiation between VT2 and VT2v was achieved using two oligonucleotide primers pairs in PCR and showed that all of 34 VT2+ porcine isolates, of which 23 were 0138:K1, harboured VT2v whereas 20 VT2+ bovine and ovine isolates harboured VT2. No isolate harboured both VT2 polymorphs. Simplified methods for sample preparation for PCR were examined and showed that PCR was not inhibited by direct addition of broth culture to the reaction mixture.
设计了寡核苷酸引物，用于特异性聚合酶链反应（PCR）扩增肠毒素STIa和LTI以及志贺毒素VT1和VT2。从猪、牛和羊腹泻病例中分离出的184株大肠杆菌，通过PCR和基因探针检测，均呈现相同的毒素谱。利用PCR中的两对寡核苷酸引物实现了VT2和VT2v的区分，结果显示，34株VT2阳性猪分离株（其中23株为0138:K1）均携带VT2v，而20株VT2阳性牛和羊分离株携带VT2。没有分离株同时携带两种VT2多态性。对PCR样品制备的简化方法进行了检测，结果表明，将肉汤培养物直接加入反应混合物中不会抑制PCR。
