Fukushima Hiroshi, Katsube Kazunori, Tsunomori Yoshie, Kishi Ryoko, Atsuta Junko, Akiba Yuko
Shimane Prefectural Institute of Public Health and Environmental Science, 582-1 Nishihamasada, Matsue City, Shimane 690-0122, Japan.
Int J Microbiol. 2009;2009:917623. doi: 10.1155/2009/917623. Epub 2009 Jun 24.
A set of four duplex SYBR Green I PCR (SG-PCR) assay combined with DNA extraction using QIAamp DNA Stool Mini kit was evaluated for the detection of foodborne bacteria from 21 foodborne outbreaks. The causative pathogens were detected in almost all cases in 2 hours or less. The first run was for the detection of 8 main foodborne pathogens in 5 stool specimens within 2 hours and the second run was for the detection of other unusual suspect pathogens within a further 45 minutes. After 2 to 4 days, the causative agents were isolated and identified. The results proved that for comprehensive and rapid molecular diagnosis in foodborne outbreaks, Duplex SG-PCR assay is not only very useful, but is also economically viable for one-step differentiation of causative pathogens in fecal specimens obtained from symptomatic patients. This then allows for effective diagnosis and management of foodborne outbreaks.
使用QIAamp DNA粪便微量提取试剂盒进行DNA提取,并结合一组四重双链SYBR Green I聚合酶链式反应(SG-PCR)检测法,对21起食源性疾病暴发事件中的食源细菌进行了检测。在几乎所有病例中,致病病原体在2小时或更短时间内被检测到。第一轮检测在2小时内对5份粪便样本中的8种主要食源性病原体进行检测,第二轮检测在另外45分钟内对其他不常见的可疑病原体进行检测。2至4天后,分离并鉴定出病原体。结果证明,对于食源性疾病暴发的全面快速分子诊断,双链SG-PCR检测法不仅非常有用,而且对于从有症状患者获取的粪便样本中致病病原体的一步鉴别在经济上也是可行的。这进而有助于对食源性疾病暴发进行有效的诊断和管理。