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通过免疫磁珠分离技术从牛粪便样本中分离出的大肠杆菌O157,采用脉冲场凝胶电泳检测其遗传多样性。

Detection of genetic diversity by pulsed-field gel electrophoresis among Escherichia coli O157 isolated from bovine faecal samples by immunomagnetic separation technique.

作者信息

Vali L, Hamouda A, Pearce M C, Knight H I, Evans J, Amyes S G B

机构信息

Molecular Chemotherapy, Centre for Infectious Diseases, University of Edinburgh, Edinburgh, UK.

出版信息

Lett Appl Microbiol. 2007 Jan;44(1):19-23. doi: 10.1111/j.1472-765X.2006.02034.x.

Abstract

AIMS

Escherichia coli O157 is considered to be one of most important human pathogens of animal origin which causes serious clinical complications. One of the most common methods to isolate E. coli O157 is the immunomagnetic separation (IMS) technique which employs specific antibodies coupled to magnetic beads to bind and extract cells from enrichment broths followed by plating onto sorbitol MacConkey agar supplemented with cefixime and potassium tellurite (CT-SMAC) plates. The aim of this study was to determine strain variation by pulsed-field gel electrophoresis (PFGE) among E. coli O157 on IMS/CT-SMAC plates.

METHODS AND RESULTS

Every suspect colony of E. coli O157 was tested following isolation by the IMS/CT-SMAC technique. From 124 colonies detected; six XbaI-PFGE profiles were identified.

CONCLUSIONS

Our results demonstrate that mixed populations of E. coli O157 with distinguishable PFGE profiles that are simultaneously present in bovine faeces can be isolated with IMS/CT-SMAC technique.

SIGNIFICANCE AND IMPACT OF THE STUDY

If the aim of the study was to analyse diversity of PFGE profiles of E. coli O157 in a faecal sample following isolation by the IMS/CT-SMAC technique, at least five colonies per sample should be analysed to detect different PFGE subtypes if present.

摘要

目的

肠出血性大肠杆菌O157被认为是最重要的动物源性人类病原体之一,可引发严重的临床并发症。分离肠出血性大肠杆菌O157最常用的方法之一是免疫磁珠分离(IMS)技术,该技术利用与磁珠偶联的特异性抗体从增菌肉汤中结合并提取细胞,随后接种到添加了头孢克肟和亚碲酸钾(CT-SMAC)的山梨醇麦康凯琼脂平板上。本研究的目的是通过脉冲场凝胶电泳(PFGE)确定IMS/CT-SMAC平板上肠出血性大肠杆菌O157菌株的变异情况。

方法与结果

采用IMS/CT-SMAC技术分离后,对每一个疑似肠出血性大肠杆菌O157菌落进行检测。在检测的124个菌落中,鉴定出6种XbaI-PFGE图谱。

结论

我们的结果表明,利用IMS/CT-SMAC技术可以从牛粪中分离出具有不同PFGE图谱的混合肠出血性大肠杆菌O157群体。

研究的意义与影响

如果研究目的是分析通过IMS/CT-SMAC技术分离后粪便样本中肠出血性大肠杆菌O157的PFGE图谱多样性,若存在不同的PFGE亚型,每个样本至少应分析5个菌落以进行检测。

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