分子生物学技术在一起食物中毒检测中的应用
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(浙江省宁波市疾病预防控制中心, 浙江 宁波 315010)

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宋启发 男 副主任技师 研究方向为细菌基因组学 E-mail:songqf@nbcdc.org.cn

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宁波市农业社会发展项目资助(2011C50041)


Application of molecular biological technology in a food poisoning incident
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(Ningbo Center for Disease Control and Prevention,Zhejiang Ningbo 315010,China)

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    摘要:

    应用分子生物学技术对一起食物中毒样品进行检测,提高实验室应对食物中毒快速检测和溯源分析能力。方法 应用实时荧光PCR技术对一起食物中毒10份患者粪便和6份可疑食品进行快速检测,并对39份粪便和8份食品进行病原培养分离鉴定,应用PCR技术对分离菌株进行invA毒力基因检测,PFGE及MLST基因分型技术对分离菌株进行同源性分析,并与其他地区菌株进行遗传学差异对比。结果 10份患者粪便和6份可疑食物经实时荧光PCR检测均为沙门菌阳性,从39份粪便和8份食品中共分离到31株肠炎沙门菌。PFGE及MLST分析显示31株菌具同源性,表明食物和患者分离菌株基因型别一致,MLST分型显示本次分离株与其他地区优势克隆有同源性,31株肠炎沙门菌均具有invA毒力基因。结论 实时荧光PCR技术应用于食物中毒病原检测,缩短了病原检测周期,提高了检测的准确性,PFGE及MLST两种基因分型技术可对病原菌进行溯源分析,对于掌握病原菌流行规律具有重要意义。

    Abstract:

    To improve the ability of rapid detection and tracing the infectious source of food poisoning outbreak with molecular biological technology.MethodsRapid detection of 10 fecal samples and 6 food samples were carried out with real-time fluorescent PCR. A total of 39 fecal samples and 8 food samples were detected by culture, isolation and identification method. Virulence gene of invA was detected by PCR. Homologous relationship among strains from local region and other regions was analyzed with PFGE and MLST genotyping technique respectively. ResultsSalmonella was detected in 10 fecal samples and 6 food samples with real-time fluorescent PCR. Thirty-one strains of Salmonella enteritidis were finally isolated and identified in 39 fecal samples and 8 food samples. Homology among those 31 strains was proved by PFGE, which suggested that the strains from food samples and clinical samples were from the same origin. Homology among the isolates and prevalent strains in other regions was also shown with MLST genotyping results. Virulence gene of invA in all isolates was testified by PCR. ConclusionTimeliness and accuracy of strain identification during food poisoning incident could been improved by real-time fluorescent PCR. Combination of PFGE and MLST genotyping techniques was of great significance to trace homologous relationship among strains in local region and other regions, and to understand their epidemiological characteristics.

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宋启发,王爱红,杨元斌,叶硕,郭延波.分子生物学技术在一起食物中毒检测中的应用[J].中国食品卫生杂志,2014,26(3):254-258.

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  • 收稿日期:2014-01-02
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  • 在线发布日期: 2014-07-16
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