Ooka Tadasuke, Terajima Jun, Kusumoto Masahiro, Iguchi Atsushi, Kurokawa Ken, Ogura Yoshitoshi, Asadulghani Md, Nakayama Keisuke, Murase Kazunori, Ohnishi Makoto, Iyoda Sunao, Watanabe Haruo, Hayashi Tetsuya
Division of Microbiology, Department of Infectious Diseases, Faculty of Medicine, University of Miyazaki, Miyazaki 889-1692, Japan.
J Clin Microbiol. 2009 Sep;47(9):2888-94. doi: 10.1128/JCM.00792-09. Epub 2009 Jul 29.
Enterohemorrhagic Escherichia coli O157 (EHEC O157) is a food-borne pathogen that has raised worldwide public health concern. The development of simple and rapid strain-typing methods is crucial for the rapid detection and surveillance of EHEC O157 outbreaks. In the present study, we developed a multiplex PCR-based strain-typing method for EHEC O157, which is based on the variability in genomic location of IS629 among EHEC O157 strains. This method is very simple, in that the procedures are completed within 2 h, the analysis can be performed without the need for special equipment or techniques (requiring only conventional PCR and agarose gel electrophoresis systems), the results can easily be transformed into digital data, and the genes for the major virulence markers of EHEC O157 (the stx(1), stx(2), and eae genes) can be detected simultaneously. Using this method, 201 EHEC O157 strains showing different XbaI digestion patterns in pulsed-field gel electrophoresis (PFGE) analysis were classified into 127 types, and outbreak-related strains showed identical or highly similar banding patterns. Although this method is less discriminatory than PFGE, it may be useful as a primary screening tool for EHEC O157 outbreaks.
肠出血性大肠杆菌O157（EHEC O157）是一种食源性病原体，已引起全球公众健康关注。开发简单快速的菌株分型方法对于EHEC O157疫情的快速检测和监测至关重要。在本研究中，我们基于EHEC O157菌株中IS629基因组位置的变异性，开发了一种基于多重PCR的EHEC O157菌株分型方法。该方法非常简单，因为程序可在2小时内完成，无需特殊设备或技术（仅需常规PCR和琼脂糖凝胶电泳系统）即可进行分析，结果可轻松转换为数字数据，并且可同时检测EHEC O157主要毒力标记基因（stx(1)、stx(2)和eae基因）。使用该方法，在脉冲场凝胶电泳（PFGE）分析中显示不同XbaI消化模式的201株EHEC O157菌株被分为127种类型，与疫情相关的菌株显示出相同或高度相似的条带模式。虽然该方法的鉴别能力不如PFGE，但它可能作为EHEC O157疫情的初步筛查工具。
