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对从波兰山羊中分离出的伪结核棒状杆菌菌株进行DNA指纹分析的三种方法的评估。

Evaluation of three methods for DNA fingerprinting of Corynebacterium pseudotuberculosis strains isolated from goats in Poland.

作者信息

Stefańska Ilona, Rzewuska Magdalena, Binek Marian

机构信息

Department of Preclinical Sciences - SGGW, Faculty of Veterinary Medicine, Warsaw University of Life Sciences, Warsaw, Poland.

出版信息

Pol J Microbiol. 2008;57(2):105-12.

Abstract

Phenotypic approaches based on metabolic and biological characteristics of Corynebacterium pseudotuberculosis have been limited due to insufficient discrimination between closely related isolates. In this paper we present performance and convenience of three molecular typing methods: BOX-PCR, random amplification of polymorphic DNA (RAPD) and amplification of DNA fragments surrounding rare restriction site (ADSRRS-fingerprinting) in genome analysis of these bacteria. Among examined 61 strains there were distinguished four, eight and 10 different genotypes by BOX-PCR, RAPD and ADSRRS-fingerprinting, respectively. The value of discrimination index was the lowest for BOX-PCR (D = 0.265), much bigger for RAPD (D = 0.539) and the highest for ADSRRS-fingerprinting (D = 0.604). The good discriminatory ability and reproducibility of RAPD and ADSRRS-fingerprinting indicates that those techniques may be particularly applied for epidemiological studies of C. pseudotuberculosis isolates. We found that ADSRRS-fingerprinting is a rapid method offering good discrimination power, excellent reproducibility and may be applied for epidemiological studies of intraspecific genetic relatedness of C. pseudotuberculosis strains.

摘要

基于伪结核棒状杆菌代谢和生物学特性的表型分析方法,由于难以有效区分密切相关的菌株而受到限制。本文介绍了三种分子分型方法在这些细菌基因组分析中的性能和便利性,即BOX-PCR、随机扩增多态性DNA(RAPD)以及围绕稀有限制位点的DNA片段扩增(ADSRRS指纹图谱)。在所检测的61株菌株中,BOX-PCR、RAPD和ADSRRS指纹图谱分别区分出4种、8种和10种不同的基因型。BOX-PCR的鉴别指数最低(D = 0.265),RAPD的鉴别指数更高(D = 0.539),ADSRRS指纹图谱的鉴别指数最高(D = 0.604)。RAPD和ADSRRS指纹图谱良好的鉴别能力和可重复性表明,这些技术尤其适用于伪结核棒状杆菌分离株的流行病学研究。我们发现,ADSRRS指纹图谱是一种快速方法,具有良好的鉴别力、出色的可重复性,可用于伪结核棒状杆菌菌株种内遗传相关性的流行病学研究。

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