Identification of Dermatophytes based on polymorphysms of nuclear and mitochondrial ribosomal DNA regions

本研究的目的是评价分子生物学技术在皮肤癣菌鉴定中的应用价值。采用限制性片段长度多态性（RFLP）分析技术，对线粒体（mtDNA）核糖体（r）DNA区域进行RFLP分析，对基因组DNA中rDNA间隔区进行RFLP分析，对基因组DNA中rDNA内转录间隔区（ITS）进行序列分析。红色毛癣菌和须根植物但当用Hae Ⅲ和Msp Ⅰ消化扩增产物时，这两个物种之间的RFLP图谱没有差异。该技术可用于T.红色毛癣菌和然而，该技术所获得的信息量相对较少，因此，对基因组DNA中rDNA的ITS 1进行序列分析具有许多优点。述的方法 ...更多信息 具有高度的可重复性和高度的灵敏度。此外，一旦确定了序列，这些数据就可以在其他地方使用，并参考储存在基因库中的数据。本研究对几种不同种类的T. mentagrophytes complex、须毛藓T. rubrum和E.絮状的ITS 1+2区的消化（约. 700 bp）。我指出这些特征是物种特异性的。该方法的灵敏度和重现性均能满足种水平鉴定的要求。该方法也适用于多个分离株的同时检测，具有成本低、检测时间短等优点，建议在实际应用中采用Mva I或Hinf I对ITS 1+2区进行PCR-RFLP分析，以鉴定皮肤癣菌。如果结果不清楚或不能接受，则确定ITS 1的核苷酸序列，并将数据提交到基因库中。少

The present study was conducted to evaluate the efficacy of molecular techniques for the identification of dermatophytes isolated from human lesions. The techniques used in the study were, restriction fragment length polymorphisms (RFLP) of ribosomal (r) DNA regions in mitochondrial (mt) DNA, RFLP analysis of spacer regions of rDNA in genomic DNA, and sequence analysis of internal transcribed spacer (ITS) of rDNA in genomic DNA.Using RFLP of PCR amplicons of rDNA region of mtDNA, the profiles generated by digestion with restriction enzyme HinA III showed differences between T. rubrum and T. mentagrophytes. But there was no difference between the RFLP profiles between these two species when the amplicons were digested with Hae III and Msp I. The technique could be used for the differenciation between T. rubrum and T. mentagrophytes, but the information obtained by the technique was rather minimum.There were many advantages of sequence analysis of ITS1 of rDNA in genomic DNA. The method … More was highly reproducible and highly sensitive. Moreover, once the sequence is determined, the data could be used elsewhere and referred to data deposited in gene banks. The expense of the method was, however, the drawback in the application in practice.PCR-RFLP analysis of ITS regions were performed on several species in the T. mentagrophytes complex, T. rubrum and E. floccosum. Digestion of ITS 1+2 regions (ca. 700 bp) by Mva I or Hint. I indicated these profiles to be species specific. The sensitivity and reproducibility was enough for the species level identification. Also the technique was inexpensive and time saving, therefore, the method is also suitable for treatment of a number of isolate at the same time.In conclusion, I propose to use PCR-RFLP analysis of ITS 1+2 regions by Mva I or Hinf I for the identification of dermatophytes in practice. If the result is unclear or unacceptable, then determine the nucleotide sequence of ITS 1, and refer the data to those deposited in a genebanks. Less