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用于检测牛粪便中大肠杆菌O157:H7的细菌培养法的敏感性

Sensitivity of bacteriologic culture for detection of Escherichia coli O157:H7 in bovine feces.

作者信息

Sanderson M W, Gay J M, Hancock D D, Gay C C, Fox L K, Besser T E

机构信息

Department of Veterinary Clinical Sciences, Washington State University, Pullman 99164, USA.

出版信息

J Clin Microbiol. 1995 Oct;33(10):2616-9. doi: 10.1128/jcm.33.10.2616-2619.1995.

Abstract

The sensitivities of several plating and broth enrichment methods for the detection of Escherichia coli O157:H7 in (i) bovine fecal samples directly inoculated with E. coli O157:H7, (ii) fecal samples from cattle in herds previously positive for E. coli O157:H7, and (iii) fecal samples from calves shedding E. coli O157:H7 after experimental oral inoculation were compared. Three enrichment protocols and three plating protocols were evaluated with directly inoculated fecal samples. All broth enrichment methods were superior to direct plating when they were combined with subsequent plating on sorbitol-MacConkey with cefixime and tellurite (SMACct). SMACct was the most sensitive plating medium, and the three alternative broth enrichment methods gave similar improvements in sensitivity. Of 351 fecal samples from known positive herds, 24 samples (6.8%) were positive by one or more methods. By the most sensitive plating method, cultures of 10-g samples were slightly more sensitive (19 of 351 [5.4%]) than cotton-tipped swab fecal samples (14 of 351 [4.0%]); however, this difference was not significant. For samples from calves orally inoculated with E. coli O157:H7, separation by immunomagnetic beads was slightly more sensitive (79%) than broth enrichment followed by plating at two dilutions (10(-3) and 10(-4)) (71%); however, this difference was not significant. The combination of overnight enrichment of swab fecal samples (0.1 g) and plating on SMACct at two dilutions (10(-3) and 10(-4)) appears to be a sensitive method for detection in large-scale studies involving hundreds of samples per week.

摘要

比较了几种平板接种和肉汤增菌方法对以下样本中大肠杆菌O157:H7的检测灵敏度:(i)直接接种大肠杆菌O157:H7的牛粪便样本;(ii)先前大肠杆菌O157:H7检测呈阳性牛群的粪便样本;(iii)实验性口服接种后排出大肠杆菌O157:H7的犊牛粪便样本。用直接接种的粪便样本评估了三种增菌方案和三种平板接种方案。当所有肉汤增菌方法与随后在含头孢克肟和亚碲酸盐的山梨醇麦康凯培养基(SMACct)上进行平板接种相结合时,均优于直接平板接种。SMACct是最灵敏的平板接种培养基,三种替代肉汤增菌方法在灵敏度提高方面效果相似。在351份已知阳性牛群的粪便样本中,有24份样本(6.8%)通过一种或多种方法检测呈阳性。通过最灵敏的平板接种方法,10克样本培养物的灵敏度略高于棉拭子粪便样本(351份样本中有19份[5.4%]对351份样本中有14份[4.0%]);然而,这种差异并不显著。对于口服接种大肠杆菌O157:H7的犊牛样本,通过免疫磁珠分离的灵敏度略高于肉汤增菌后在两种稀释度(10⁻³和10⁻⁴)下进行平板接种(71%);然而,这种差异并不显著。在每周涉及数百份样本的大规模研究中,对拭子粪便样本(0.1克)进行过夜增菌并在两种稀释度(10⁻³和10⁻⁴)下在SMACct上进行平板接种的组合似乎是一种灵敏的检测方法。

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