Korabecná M, Liska V, Fajfrlík K
Institute for Biology, Faculty of Medicine, Charles University, 301 66 Pilsen, Czechia.
Folia Microbiol (Praha). 2003;48(2):233-8. doi: 10.1007/BF02930961.
Restriction fragment length polymorphism analysis of the 5.8S rRNA gene and the internal transcribed spacers (ITS1 and ITS2) was used for examination of 66 isolates belonging to 19 species. Intraspecies variability was found in the examined region of 11 species (Candida albicans, C. catenulata, C. colliculosa, C. glabrata, C. kefyr, C. melinii, C. parapsilosis, C. guillermondii, C. solanii, C. tropicalis, Saccharomyces cerevisiae). Region of ITS-5.8S rDNA was amplified using the primers ITS1 and ITS4. The amplicons were digested by HaeIII, HinfI and CfoI. The recognized intraspecies variability was confirmed in the second step, in which the shorter fragments of this region were amplified using primers ITS1 and ITS2 and analyzed by capillary electrophoresis. Considerable intraspecific variability renders this method unsuitable for species identification, whereas it can be useful for epidemiological tracing of isolates.
利用5.8S rRNA基因及内部转录间隔区（ITS1和ITS2）的限制性片段长度多态性分析，对属于19个种的66株分离株进行检测。在所检测的11个种（白色念珠菌、链状念珠菌、小巢状念珠菌、光滑念珠菌、克菲念珠菌、梅氏念珠菌、近平滑念珠菌、季也蒙念珠菌、索拉尼念珠菌、热带念珠菌、酿酒酵母）的区域发现了种内变异性。使用引物ITS1和ITS4扩增ITS - 5.8S rDNA区域。扩增产物用HaeIII、HinfI和CfoI进行酶切。第二步证实了所识别的种内变异性，其中使用引物ITS1和ITS2扩增该区域的较短片段，并通过毛细管电泳进行分析。种内存在相当大的变异性，使得该方法不适用于菌种鉴定，然而它可用于分离株的流行病学追踪。
