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使用(GTG)₅-PCR 指纹图谱鉴定葡萄球菌属。

Identification of Staphylococcus spp. using (GTG)₅-PCR fingerprinting.

机构信息

Czech Collection of Microorganisms, Department of Experimental Biology, Faculty of Science, Masaryk University, Tvrdého 14, 602 00, Brno, Czech Republic.

出版信息

Syst Appl Microbiol. 2010 Dec;33(8):451-6. doi: 10.1016/j.syapm.2010.09.004. Epub 2010 Nov 20.

Abstract

A group of 212 type and reference strains deposited in the Czech Collection of Microorganisms (Brno, Czech Republic) and covering 41 Staphylococcus species comprising 21 subspecies was characterised using rep-PCR fingerprinting with the (GTG)₅ primer in order to evaluate this method for identification of staphylococci. All strains were typeable using the (GTG)₅ primer and generated PCR products ranging from 200 to 4500 bp. Numerical analysis of the obtained fingerprints revealed (sub)species-specific clustering corresponding with the taxonomic position of analysed strains. Taxonomic position of selected strains representing the (sub)species that were distributed over multiple rep-PCR clusters was verified and confirmed by the partial rpoB gene sequencing. Staphylococcus caprae, Staphylococcus equorum, Staphylococcus sciuri, Staphylococcus piscifermentans, Staphylococcus xylosus, and Staphylococcus saprophyticus revealed heterogeneous fingerprints and each (sub)species was distributed over several clusters. However, representatives of the remaining Staphylococcus spp. were clearly separated in single (sub)species-specific clusters. These results showed rep-PCR with the (GTG)₅ primer as a fast and reliable method applicable for differentiation and straightforward identification of majority of Staphylococcus spp.

摘要

一组 212 株类型和参考菌株存放在捷克微生物收藏(布尔诺,捷克共和国)中,涵盖了 41 种葡萄球菌属物种,包括 21 个亚种,使用(GTG)₅引物的重复聚合酶链反应指纹图谱进行了特征描述,以评估该方法对葡萄球菌的鉴定能力。所有菌株均可用(GTG)₅引物进行分型,产生的 PCR 产物大小在 200 到 4500bp 之间。所得指纹图谱的数值分析显示出与分析菌株分类地位相对应的(亚种)特异性聚类。通过部分 rpoB 基因测序,验证并确认了代表分布在多个重复聚合酶链反应聚类中的(亚种)的选定菌株的分类地位。山羊葡萄球菌、马胃葡萄球菌、松鼠葡萄球菌、鱼精蛋白发酵葡萄球菌、木糖葡萄球菌和腐生葡萄球菌显示出异质的指纹图谱,每个(亚种)都分布在几个聚类中。然而,其余葡萄球菌属的代表则明显分离在单一的(亚种)特异性聚类中。这些结果表明,(GTG)₅引物的重复聚合酶链反应是一种快速可靠的方法,适用于大多数葡萄球菌属的分化和直接鉴定。

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