弥散黏附性大肠埃希菌多重PCR检测方法的建立及其在感染性腹泻患者中的流行情况

阳波 张网 郭秋生 白向宁 许彦梅 熊衍文 罗成旺

阳波, 张网, 郭秋生, 白向宁, 许彦梅, 熊衍文, 罗成旺. 弥散黏附性大肠埃希菌多重PCR检测方法的建立及其在感染性腹泻患者中的流行情况[J]. 疾病监测, 2017, 32(5): 423-427. doi: 10.3784/j.issn.1003-9961.2017.05.017
引用本文: 阳波, 张网, 郭秋生, 白向宁, 许彦梅, 熊衍文, 罗成旺. 弥散黏附性大肠埃希菌多重PCR检测方法的建立及其在感染性腹泻患者中的流行情况[J]. 疾病监测, 2017, 32(5): 423-427. doi: 10.3784/j.issn.1003-9961.2017.05.017
YANG Bo, ZHANG Wang, GUO Qiu-sheng, BAI Xiang-ning, XU Yan-mei, XIONG Yan-wen, LUO Cheng-wang. Establishment of multiplex polymerase chain reaction assay for detection of diffuse adherent Escherichia coli in diarrhea patients[J]. Disease Surveillance, 2017, 32(5): 423-427. doi: 10.3784/j.issn.1003-9961.2017.05.017
Citation: YANG Bo, ZHANG Wang, GUO Qiu-sheng, BAI Xiang-ning, XU Yan-mei, XIONG Yan-wen, LUO Cheng-wang. Establishment of multiplex polymerase chain reaction assay for detection of diffuse adherent Escherichia coli in diarrhea patients[J]. Disease Surveillance, 2017, 32(5): 423-427. doi: 10.3784/j.issn.1003-9961.2017.05.017

弥散黏附性大肠埃希菌多重PCR检测方法的建立及其在感染性腹泻患者中的流行情况

doi: 10.3784/j.issn.1003-9961.2017.05.017
基金项目: 

国家自然科学基金(No.81371762)

Establishment of multiplex polymerase chain reaction assay for detection of diffuse adherent Escherichia coli in diarrhea patients

Funds: 

This study was supported by the National Natural Science Foundation of China (No. 81371762)

  • 摘要: 目的 建立一种快速检测弥散黏附性大肠埃希菌(DAEC)的普通多重PCR方法,了解DAEC在腹泻患者中的流行情况。方法 根据DAEC黏附基因afaB、afaC、afaD、daaE及16S rRNA基因rrs,建立4重PCR反应体系,对PCR引物、反应体系和反应条件进行优化,评价敏感性和特异性,并应用于389份腹泻患者粪便标本的筛查。结果 4重PCR反应可以特异性扩增DAEC菌株afaB/C、afaD、daaE基因片段,除2株肠集聚性大肠埃希菌(EAEC)外,检测引物对其他致泻性大肠埃希菌及常见肠道病原菌均无特异性扩增,此多重PCR方法的检测下限可达3.20 101 CFU/反应(8.01 103 CFU/ml)。利用本研究建立的多重PCR方法对腹泻患者粪便标本进行检测,发现DAEC菌株分离率为6.2%(24/389)。结论 本研究建立的多重PCR方法可用于DAEC菌株的快速鉴别,也可用于人粪便标本DAEC的初步筛查。
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出版历程
  • 收稿日期:  2016-11-17
  • 刊出日期:  2017-05-20

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