刘晓琳, 王慧, 郭凯, 曲剑英, 王丽娟, 于维森. 联合分子检测法对腹泻患者粪便中致泻性大肠埃希菌的调查[J]. 疾病监测, 2020, 35(12): 1110-1114. DOI: 10.3784/j.issn.1003-9961.2020.12.011
引用本文: 刘晓琳, 王慧, 郭凯, 曲剑英, 王丽娟, 于维森. 联合分子检测法对腹泻患者粪便中致泻性大肠埃希菌的调查[J]. 疾病监测, 2020, 35(12): 1110-1114. DOI: 10.3784/j.issn.1003-9961.2020.12.011
Liu Xiaolin, Wang Hui, Guo Kai, Qu Jianying, Wang Lijuan, Yu Weisen. Detection of diarrheagenic Escherichia coli from diarrhea cases’ stool samples with combined molecular detection method[J]. Disease Surveillance, 2020, 35(12): 1110-1114. DOI: 10.3784/j.issn.1003-9961.2020.12.011
Citation: Liu Xiaolin, Wang Hui, Guo Kai, Qu Jianying, Wang Lijuan, Yu Weisen. Detection of diarrheagenic Escherichia coli from diarrhea cases’ stool samples with combined molecular detection method[J]. Disease Surveillance, 2020, 35(12): 1110-1114. DOI: 10.3784/j.issn.1003-9961.2020.12.011

联合分子检测法对腹泻患者粪便中致泻性大肠埃希菌的调查

Detection of diarrheagenic Escherichia coli from diarrhea cases’ stool samples with combined molecular detection method

  • 摘要:
      目的  采用胃肠道感染测试条(FA GI)联合实时聚合酶链式反应(PCR)直接检测法,回顾性调查山东省青岛市腹泻患者致泻性大肠埃希菌(DEC)的感染情况及菌株基因型,并与传统方法进行比较,探讨不同检测方法在腹泻监测中的应用。
      方法  对2018年青岛市2家哨点医院采集的263份腹泻粪便标本,采用传统培养法分离大肠埃希菌并送至青岛市疾病预防控制中心,利用多重PCR方法鉴定DEC;同时采用FA GI联合实时PCR的方法直接检测粪便中的DEC。 将每5份粪便样标本混合为1份混合样品,用FA GI进行检测,再用实时PCR方法检测阳性混合样品中的单个样品。
      结果  263份标本中,采用传统分离培养联合多重PCR法检测,15份样品阳性,阳性率为5.7%。 其中,肠产毒性大肠埃希菌(ETEC)、肠致病性大肠埃希菌(EPEC)、肠集聚性大肠埃希菌(EAEC)、肠侵袭性大肠埃希菌(EIEC)阳性的标本分别为7、4、3、1份。 采用FA GI联合实时PCR直接检测法检测,阳性样品35 份,阳性率为13.3%。 其中ETEC、EAEC、EPEC、EIEC阳性的标本分别为12、12、9、2份。 2种方法均未检到产志贺毒素大肠埃希菌(STEC)阳性的标本。 2种方法毒力基因检测结果一致。 EAEC、ETEC、EPEC携带的主要毒力基因分别为picestIbeae
      结论  FA GI联合实时PCR直接检测法显著提高了粪便中DEC的检出率,敏感性和特异性高,省时省力,可以辅助DEC常规监测,准确地反映DEC的流行和分布情况,为DEC的防控提供有力的实验室依据。

     

    Abstract:
      Objective  To retrospective investigate the infection status and genotype of diarrheagenic Escherichia coli(DEC)in diarrhea cases in Qingdao, Shandong with FilmArray gastrointestinal panel (FA GI) combining with real-time PCR, and the results were compared with traditional detection method to discuss the application of different detection method in diarrhea surveillance.
      Methods  A total of 263 stool samples were collected from diarrhea cases by two hospitals in Qingdao. DEC strains were isolated by traditional culture method and submitted to Qingdao Municipal Center for Disease Control and Prevention for the identification by multiple real-time PCR, and at the same time DEC was detected directly from stool samples by FA GI combining with real-time PCR. Every five stool samples were mixed for one detection by FA GI. The positive results indicated that every sample needed to be detected again for DEC by real-time PCR.
      Results  A total of 15 samples were DEC positive (5.7%) by traditional culture, in which 7, 4, 3, 1 were ETEC, EPEC, EAEC, EIEC positive, respectively. 35 DEC positive samples (13.3%) were detected with FA GI, in which 12, 12, 9, 2 were ETEC, EAEC, EPEC and EIEC positive, respectively. No sample was Shiga toxin-producing Escherichia coli (STEC) positive by both two methods. The virulence genes carriages in DEC indicated by two different methods were similar, i.e. pic in EAEC, estIb in ETEC and eae in EPEC.
      Conclusion  The direct detection method of FA GI combining with real-time PCR can significantly improve the positive detection rate of DEC in stool samples from diarrhea cases due to its high sensitivity and specificity. This detection method can save both time and labor and can be used as supplementary investigation for routine surveillance for the better understanding of the prevalence and genotype distribution of DEC in diarrhea patients and strengthening of laboratory capability.

     

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