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通过DNA定量开发一种快速可靠的方法来评估纺织品上的微生物定植和生长情况。

Development of a fast and reliable method for the assessment of microbial colonization and growth on textiles by DNA quantification.

作者信息

Teufel Linda, Schuster K Christian, Merschak Petra, Bechtold Thomas, Redl Bernhard

机构信息

Christian Doppler Laboratory for Textile and Fibre Chemistry in Cellulosics, Institute of Textile Chemistry and Textile Physics, Leopold-Franzens University Innsbruck, Dornbirn, Austria.

出版信息

J Mol Microbiol Biotechnol. 2008;14(4):193-200. doi: 10.1159/000108657. Epub 2007 Sep 20.

Abstract

There is a lack of relevant methods to assess the colonization of textiles by skin bacteria because present methods are mainly culture-based procedures. Therefore, the goal of this study was to develop a fast and sensitive culture-independent procedure for the quantification of microbial colonization and growth on textiles. We have established a suitable protocol to use DNA quantification as a reliable method for in vitroand in vivoinvestigations of textiles. For DNA extraction, a two-step procedure comprising treatment of the textile with a solution containing Triton X-100 and lysozyme for 1 h and a successive treatment by SDS and proteinase K for 2 h turned out to be most efficient. DNA extracted from textiles and fabrics was than quantified with the highly sensitive PicoGreen fluorescent dye. In vitrochallenge tests demonstrated a strong correlation between numbers of bacteria on textiles and amount of DNA extracted from textiles. Therefore, this method was used to compare different materials after in vivotrials for assessment of their susceptibility for microbial colonization and growth.

摘要

由于目前的方法主要是基于培养的程序,因此缺乏评估皮肤细菌在纺织品上定殖的相关方法。因此,本研究的目标是开发一种快速、灵敏的非培养程序,用于定量纺织品上的微生物定殖和生长。我们已经建立了一个合适的方案,将DNA定量作为一种可靠的方法用于纺织品的体外和体内研究。对于DNA提取,包括用含有Triton X-100和溶菌酶的溶液处理纺织品1小时,然后依次用SDS和蛋白酶K处理2小时的两步程序被证明是最有效的。从纺织品和织物中提取的DNA随后用高灵敏度的PicoGreen荧光染料进行定量。体外挑战试验表明,纺织品上的细菌数量与从纺织品中提取的DNA量之间存在很强的相关性。因此,该方法用于体内试验后比较不同材料,以评估它们对微生物定殖和生长的敏感性。

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