New 16-plex PCR method for rapid detection of diarrheagenic Escherichia coli directly from stool samples
New 16-plex PCR method for rapid detection of diarrheagenic Escherichia coli directly from stool samples
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DOI：
10.1007/s10096-009-0720-x
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发表时间：
2009-08-01
影响因子：
4.5
通讯作者：
Kirveskari, J.
中科院分区：
文献类型：
--
作者：
Antikainen, J.;Tarkka, E.;Kirveskari, J.
建立了一种适用于大肠埃希菌（EHEC、EIEC、EAEC、ETEC和EPEC）常规诊断的快速16重聚合酶链反应（PCR），并用对照菌株进行了验证，并对250份粪便样品进行了检测。当用289个对照细菌菌株进行测试时，特异性为100%，并且通过煮沸细菌培养物（10（4）-10（5）菌落形成单位/ml）来获得直接从粪便样品自动DNA提取的分析灵敏度。直接从粪便DNA提取开始的测定设计允许当天分析，而不损害灵敏度和特异性，这使得其与培养细菌后的PCR相比具有上级优势。16重PCR法检测大肠埃希氏菌感染率高。从国外返回者粪便标本中大肠杆菌阳性率为39.0%，无出国史者粪便标本中大肠杆菌阳性率为8.7%（P < 0.001）。大肠埃希氏菌的高流行率。大肠杆菌的研究表明，他们的筛查应该是正常腹泻诊断的一部分，至少在领先的诊断实验室。
A rapid 16-plex polymerase chain reaction (PCR) suitable for routine diagnostics of diarrheagenic Escherichia coli (EHEC, EIEC, EAEC, ETEC, and EPEC) was developed, validated with control strains, and tested with 250 diarrhoeal stool samples. The specificity was 100% when tested with 289 control bacterial strains, and the analytical sensitivity of automated DNA extraction directly from stool samples was made by boiling the bacterial culture (10(4)-10(5) colony forming units/ml). The assay design starting directly from extraction of stool DNA allowed same day analysis without compromising sensitivity and specificity, which makes it superior compared to PCR after culturing the bacteria. The 16-plex PCR method demonstrated high prevalence of diarrheagenic E. coli in stool samples of patients returning from abroad (39.0%) in contrast to the patients with no travel history (8.7%; p < 0.001). The high prevalence of diarrheagenic E. coli suggests that their screening should be part of normal diarrhoea diagnostics, at least in the leading diagnostic laboratories.