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多种基因座可变串联重复序列分析在追踪食品加工环境中金黄色葡萄球菌的有效应用。

Effective application of multiple locus variable number of tandem repeats analysis to tracing Staphylococcus aureus in food-processing environment.

机构信息

Department of Microbiology and Molecular Biology, Food Research Institute, Bratislava, Slovakia.

出版信息

Lett Appl Microbiol. 2014 Apr;58(4):376-83. doi: 10.1111/lam.12200. Epub 2013 Dec 11.

Abstract

UNLABELLED

A total of 256 isolates of Staphylococcus aureus were isolated from 98 samples (34 swabs and 64 food samples) obtained from small or medium meat- and cheese-processing plants in Slovakia. The strains were genotypically characterized by multiple locus variable number of tandem repeats analysis (MLVA), involving multiplex polymerase chain reaction (PCR) with subsequent separation of the amplified DNA fragments by an automated flow-through gel electrophoresis. With the panel of isolates, MLVA produced 31 profile types, which was a sufficient discrimination to facilitate the description of spatial and temporal aspects of contamination. Further data on MLVA discrimination were obtained by typing a subpanel of strains by multiple locus sequence typing (MLST). MLVA coupled to automated electrophoresis proved to be an effective, comparatively fast and inexpensive method for tracing S. aureus contamination of food-processing factories.

SIGNIFICANCE AND IMPACT OF THE STUDY

Subspecies genotyping of microbial contaminants in food-processing factories may facilitate identification of spatial and temporal aspects of the contamination. This may help to properly manage the process hygiene. With S. aureus, multiple locus variable number of tandem repeats analysis (MLVA) proved to be an effective method for the purpose, being sufficiently discriminative, yet comparatively fast and inexpensive. The application of automated flow-through gel electrophoresis to separation of DNA fragments produced by multiplex PCR helped to improve the accuracy and speed of the method.

摘要

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从斯洛伐克的小型和中型肉类和奶酪加工厂采集的 98 个样本(34 个拭子和 64 个食品样本)中分离出了 256 株金黄色葡萄球菌。通过多位点可变数目串联重复分析(MLVA)对菌株进行了基因分型特征分析,涉及多重聚合酶链反应(PCR),随后通过自动电泳分离扩增的 DNA 片段。使用该组分离株,MLVA 产生了 31 种图谱类型,足以促进污染的时空方面的描述。通过对亚组菌株进行多位点序列分型(MLST)对 MLVA 区分度进行了进一步的数据。MLVA 与自动电泳相结合被证明是一种有效的、相对快速且廉价的方法,可用于追踪食品加工厂中金黄色葡萄球菌的污染。

研究的意义和影响

对食品加工厂中微生物污染物的亚种进行基因分型可能有助于识别污染的时空方面。这有助于正确管理工艺卫生。对于金黄色葡萄球菌,多位点可变数目串联重复分析(MLVA)被证明是一种有效的方法,因为它具有足够的区分度,但相对快速且廉价。将自动电泳用于分离多重 PCR 产生的 DNA 片段有助于提高该方法的准确性和速度。

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