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用于肠炎沙门氏菌亚种分型的通用多位点可变数目串联重复序列分析与脉冲场凝胶电泳的开发及比较

Development and comparison of a generic multiple-locus variable-number tandem repeat analysis with pulsed-field gel electrophoresis for typing of Salmonella enterica subsp. enterica.

作者信息

Kjeldsen M K, Torpdahl M, Pedersen K, Nielsen E M

机构信息

The Unit for Foodborne Infections, Department of Microbiology and Infection Control, Statens Serum Institut, Copenhagen, Denmark.

National Food Institute, Technical University of Denmark, Søborg, Denmark.

出版信息

J Appl Microbiol. 2015 Dec;119(6):1707-17. doi: 10.1111/jam.12965.

Abstract

AIMS

Salmonella enterica subsp. enterica causes salmonellosis in humans and animals. Serovar-specific multiple-locus variable-number tandem repeat analysis (MLVA) is widely used for Salmonella surveillance; however, isolates have to be serotyped prior to MLVA typing and only the most common serovars can be typed. We developed a MLVA scheme for high-discriminatory typing of Salmonella.

METHODS AND RESULTS

Sixty-six unique VNTRs were investigated and the polymorphisms of seven promising VNTRs were evaluated with a panel 163 diverse isolates of 14 serotypes of significance for human health. Five VNTRs were selected for MLVA analysis. The discriminatory power was evaluated within serovars by 163 isolates and MLVA yielded 79 genotypes (DI of 0·9790) and pulsed-field gel electrophoresis (PFGE) revealed 87 genotypes (DI of 0·9989). MLVA divided each serotype into 2-8 different profiles and identified six pairs of outbreak-related strains.

CONCLUSIONS

The technique showed a high-discriminatory power within most serotypes comparable with or better than that of PFGE.

SIGNIFICANCE AND IMPACT OF THE STUDY

This MLVA assay makes it possible to use a single typing method for Salmonella surveillance and outbreak investigations. This allows inexpensive and fast surveillance for laboratories without resources for both serotyping and molecular typing, e.g. PFGE or sequence-based methods, and thereby improve the effectiveness of epidemiological investigations of Salmonella infections globally.

摘要

目的

肠炎沙门氏菌亚种肠炎型可导致人和动物感染沙门氏菌病。血清型特异性多位点可变数目串联重复序列分析(MLVA)广泛用于沙门氏菌监测;然而,在进行MLVA分型之前必须先对分离株进行血清分型,且只能对最常见的血清型进行分型。我们开发了一种用于沙门氏菌高分辨率分型的MLVA方案。

方法与结果

研究了66个独特的VNTR,并使用一组163株来自14种对人类健康具有重要意义的血清型的不同分离株评估了7个有前景的VNTR的多态性。选择了5个VNTR进行MLVA分析。通过163株分离株评估了血清型内的鉴别力,MLVA产生了79种基因型(鉴别指数为0·9790),脉冲场凝胶电泳(PFGE)显示了87种基因型(鉴别指数为0·9989)。MLVA将每种血清型分为2 - 8种不同的谱型,并鉴定出6对与暴发相关的菌株。

结论

该技术在大多数血清型中显示出高鉴别力,与PFGE相当或更好。

研究的意义和影响

这种MLVA检测方法使得可以使用单一分型方法进行沙门氏菌监测和暴发调查。这使得没有资源同时进行血清分型和分子分型(如PFGE或基于序列的方法)的实验室能够进行廉价且快速的监测,从而提高全球沙门氏菌感染流行病学调查的有效性。

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