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通过聚合酶链反应检测粪便标本中的产肠毒素大肠杆菌

Detection of enterotoxigenic Escherichia coli in stool specimens by polymerase chain reaction.

作者信息

Yavzori M, Porath N, Ochana O, Dagan R, Orni-Wasserlauf R, Cohen D

机构信息

Medical Corps, Israel Defence Force, Medical School, Ben-Gurion University, Israel.

出版信息

Diagn Microbiol Infect Dis. 1998 Aug;31(4):503-9. doi: 10.1016/s0732-8893(98)00040-6.

Abstract

A polymerase chain reaction (PCR) protocol for rapid (7 h) detection of enterotoxigenic Escherichia coli (ETEC) is described. This protocol has been validated on 57 stool samples from young children by comparing it with the colony hybridization technique. A good agreement was found between the two methods with Cohen's kappa statistics of 0.87 and 0.79 for the detection of the heat-stable toxin (ST) and heat-labile toxin (LT), respectively. Of 26 samples positive for LT and 15 samples positive for ST by colony hybridization, 21 (81%) and 15 (100%) were also found to be positive for LT and ST by PCR, respectively. Only one sample identified as LT-negative by colony hybridization was found to be positive by PCR. However, 3 of 42 samples of ST-negative by colony hybridization were detected as positive by PCR. A reconstruction experiment revealed that PCR could detect LT-producing and ST-producing ETEC at minimal concentrations of 2.5 x 10(3) cfu and 2.5 x 10(2) cfu per gram of feces, respectively. These data indicate the possible use of this method for rapid identification of ETEC-associated diarrhea in clinical and epidemiological settings.

摘要

本文描述了一种用于快速(7小时)检测产肠毒素大肠杆菌(ETEC)的聚合酶链反应(PCR)方案。通过与菌落杂交技术比较,该方案已在57份幼儿粪便样本上得到验证。两种方法之间具有良好的一致性,检测耐热毒素(ST)和不耐热毒素(LT)时,Cohen's kappa统计量分别为0.87和0.79。在通过菌落杂交检测LT呈阳性的26份样本和ST呈阳性的15份样本中,通过PCR检测发现LT和ST也分别有21份(81%)和15份(100%)呈阳性。通过菌落杂交鉴定为LT阴性的样本中,只有一份通过PCR检测呈阳性。然而,通过菌落杂交检测为ST阴性的42份样本中有3份通过PCR检测呈阳性。一项重建实验表明,PCR分别在每克粪便中最低浓度为2.5×10³ cfu和2.5×10² cfu时,能够检测出产生LT和产生ST的ETEC。这些数据表明该方法可能用于临床和流行病学环境中快速鉴定与ETEC相关的腹泻。

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