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用于检测人粪便标本中产志贺毒素大肠杆菌的志贺毒素酶联免疫吸附测定法与两种聚合酶链反应的比较

Comparison of a shiga toxin enzyme-linked immunosorbent assay and two types of PCR for detection of shiga toxin-producing Escherichia coli in human stool specimens.

作者信息

Pulz Matthias, Matussek Andreas, Monazahian Masyar, Tittel Andreas, Nikolic Elisabet, Hartmann Maike, Bellin Tobias, Buer Jan, Gunzer Florian

机构信息

Niedersächsisches Landesgesundheitsamt, 30449 Hannover, Germany.

出版信息

J Clin Microbiol. 2003 Oct;41(10):4671-5. doi: 10.1128/JCM.41.10.4671-4675.2003.

Abstract

Shiga toxin (Stx)-producing Escherichia coli (STEC) is a major cause of sporadic cases of disease as well as serious outbreaks worldwide. The spectrum of illnesses includes mild nonbloody diarrhea, hemorrhagic colitis, and hemolytic-uremic syndrome. STEC produces one or more Stxs, which are subdivided into two major classes, Stx1 and Stx2. The ingestion of contaminated food or water, person-to-person spread, and contact with animals are the major transmission modes. The infective dose of STEC may be less than 100 organisms. Effective prevention of infection is dependent on rapid detection of the causative bacterial pathogen. In the present study, we examined 295 stool specimens for the presence of Stx-producing E. coli by three different methods: an Stx enzyme-linked immunosorbent assay, a conventional PCR assay, and a LightCycler PCR (LC-PCR) assay protocol recently developed by our laboratory at the Institute of Medical Microbiology at Hannover Medical School. Our intent was to compare these three methods and to examine the utility of the STEC LC-PCR protocol in a clinical laboratory. The addition of a control DNA to each sample to clearly discriminate inhibited specimens from negative ones enhanced the accuracy of the LC-PCR protocol. From our results, it can be concluded that LC-PCR is a very useful tool for the rapid and safe detection of STEC in clinical samples.

摘要

产志贺毒素大肠杆菌(STEC)是全球散发病例以及严重疫情的主要病因。疾病谱包括轻度非血性腹泻、出血性结肠炎和溶血尿毒综合征。STEC产生一种或多种志贺毒素(Stxs),这些毒素可分为两大类,即Stx1和Stx2。摄入受污染的食物或水、人传人传播以及与动物接触是主要传播方式。STEC的感染剂量可能少于100个菌体。有效预防感染取决于对致病细菌病原体的快速检测。在本研究中,我们采用三种不同方法检测了295份粪便标本中是否存在产志贺毒素大肠杆菌:一种志贺毒素酶联免疫吸附测定法、一种常规聚合酶链反应(PCR)测定法以及我们汉诺威医学院医学微生物研究所实验室最近开发的一种LightCycler PCR(LC-PCR)测定法方案。我们的目的是比较这三种方法,并检验STEC的LC-PCR方案在临床实验室中的实用性。向每个样本中添加对照DNA以明确区分受抑制标本和阴性标本,提高了LC-PCR方案的准确性。从我们的结果可以得出结论,LC-PCR是临床样本中快速、安全检测STEC的非常有用的工具。

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