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基于毛细管凝胶电泳的聚合酶链反应用于检测高毒力核糖型菌株的效用。

Usefulness of Capillary Gel Electrophoresis-Based PCR for Detection of Strains with Hypervirulent Ribotypes.

作者信息

Bogiel Tomasz, Dura Alicja, Woźniak Marcin, Mikucka Agnieszka, Kanarek Piotr

机构信息

Department of Microbiology, Ludwik Rydygier Collegium Medicum in Bydgoszcz, Nicolaus Copernicus University in Toruń, 85-094 Bydgoszcz, Poland.

Department of Clinical Microbiology, Antoni Jurasz University Hospital No. 1, 85-094 Bydgoszcz, Poland.

出版信息

Gels. 2024 May 17;10(5):343. doi: 10.3390/gels10050343.

Abstract

is a complex of anaerobic bacteria responsible for the epidemics of post-antibiotic diarrhea as one of the examples of CDI ( infection). As many as 70% of cases concern hospitalized patients, particularly those in intensive care units. Ribotyping is one of the most common methods for differentiating bacterial strains. The purpose of this work was to show the effectiveness of the gel electrophoresis-based PCR ribotyping method and the Webribo database for typing isolates, including the hypervirulent 027 ribotype. DNA samples extracted from 69 strains with previously marked genotypes were included in this study. PCR was performed using 16S-23S primers, and capillary gel electrophoresis was performed on the Applied Biosystem 3130xl Genetic Analyzer. The Webribo database was applied for ribotype assignment. Out of 69 samples, 48 belonged to already known ribotypes, 13 represented new ribotypes and 8 was indicated as similar to the existing ones, having some differences. Capillary gel electrophoresis-based PCR is an effective method for the differentiation of ribotypes and can be recognized as a very useful tool in epidemiological studies, while the Webribo database is a useful and an accessible database for a quick analysis of ribotypes.

摘要

是一种厌氧菌复合体,作为艰难梭菌感染(CDI)的例子之一,它是导致抗生素相关性腹泻流行的原因。多达70%的病例涉及住院患者,尤其是重症监护病房的患者。核糖体分型是区分细菌菌株最常用的方法之一。这项工作的目的是展示基于凝胶电泳的PCR核糖体分型方法和Webribo数据库在对分离株进行分型方面的有效性,包括高毒力的027核糖体分型。本研究纳入了从69株具有先前标记基因型的菌株中提取的DNA样本。使用16S - 23S引物进行PCR,并在Applied Biosystem 3130xl遗传分析仪上进行毛细管凝胶电泳。使用Webribo数据库进行核糖体分型的指定。在69个样本中,48个属于已知的核糖体分型,13个代表新的核糖体分型,8个被表明与现有分型相似,但存在一些差异。基于毛细管凝胶电泳的PCR是区分艰难梭菌核糖体分型的有效方法,可被认为是流行病学研究中非常有用的工具,而Webribo数据库是一个用于快速分析艰难梭菌核糖体分型的有用且可访问的数据库。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/45f3/11121280/eac41824aa90/gels-10-00343-g001.jpg

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