Lin Z, Kurazono H, Yamasaki S, Takeda Y
Department of Microbiology, Faculty of Medicine, Kyoto University, Japan.
Microbiol Immunol. 1993;37(7):543-8. doi: 10.1111/j.1348-0421.1993.tb01675.x.
We constructed common primers for the polymerase chain reaction to detect the genes for various Verotoxins reported, that is, VT1 (or SLT-I), VT2 (or SLT-II), VT2vha, VT2vhb, SLT-IIv (or VT2vp1, VTe) and SLT-IIva (or VT2vp2). A total of 80 Verocytotoxin-producing Escherichia coli strains isolated from humans, domestic animals and meats gave a positive result by PCR with the designed common primers. Digestion by restriction endonucleases BglII and EcoT14I of the amplicon of the VT2vp2 gene gave specific bands of the expected sizes, but not of the amplicons of other VT genes, suggesting a possible method for identification of the VT2vp2 gene. Application of the PCR with the designed primers in diagnostic and epidemiological studies on VTEC infection is also discussed.
我们构建了用于聚合酶链反应的通用引物,以检测已报道的各种志贺毒素基因,即VT1(或SLT-I)、VT2(或SLT-II)、VT2vha、VT2vhb、SLT-IIv(或VT2vp1、VTe)和SLT-IIva(或VT2vp2)。从人类、家畜和肉类中分离出的总共80株产志贺毒素大肠杆菌菌株,经PCR检测,所设计的通用引物呈阳性结果。用限制性内切酶BglII和EcoT14I消化VT2vp2基因的扩增子,得到预期大小的特异性条带,但其他VT基因的扩增子未出现该条带,这表明这可能是一种鉴定VT2vp2基因的方法。还讨论了所设计引物的PCR在VTEC感染诊断和流行病学研究中的应用。