Scherer S, Stevens D A
J Clin Microbiol. 1987 Apr;25(4):675-9. doi: 10.1128/jcm.25.4.675-679.1987.
Methods are described for extraction of DNA from the yeast form of Candida spp., followed by digestion and electrophoresis of DNA fragments. The resulting gel patterns (greater than 100 bands) were used to type Candida isolates. Four intense bands identified, three of which are present in each isolate (6 to 7, 3.7 or 4.2, and 2.5 to 3 kilobases), appear to be DNA encoding the rRNA. The methods proved to be both simple and reproducible. The patterns were shown to be stable through several hundred doublings from multiple single colonies. A survey of isolates showed that, on the basis of similarity of gel patterns, several Candida species could be sorted into mutually exclusive groups, and subgroups could be created. Analyses of this survey suggested the possible epidemiologic and taxonomic applications of these methods. DNA typing methods appear to offer important potential advantages over phenotyping methods. The methods provide a base for further epidemiologic studies and for further development of techniques, such as the use of cloned probes for studies of DNA homology.
本文描述了从念珠菌属酵母形态中提取DNA的方法,随后对DNA片段进行消化和电泳。所得的凝胶图谱(超过100条带)用于对念珠菌分离株进行分型。鉴定出四条密集条带,其中三条存在于每个分离株中(6至7千碱基、3.7或4.2千碱基以及2.5至3千碱基),这些条带似乎是编码rRNA的DNA。这些方法被证明既简单又可重复。图谱在来自多个单菌落的数百次传代培养中显示稳定。对分离株的一项调查表明,基于凝胶图谱的相似性,几种念珠菌物种可被分为相互排斥的组,并可进一步细分。对该调查的分析表明了这些方法在流行病学和分类学上的潜在应用。与表型分析方法相比,DNA分型方法似乎具有重要的潜在优势。这些方法为进一步的流行病学研究以及技术的进一步发展奠定了基础,例如使用克隆探针研究DNA同源性。