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

基于核和线粒体核糖体 DNA 区域多态性的皮肤癣菌鉴定

• 批准号: 10670811
• 负责人: MOCHIZUKI Takashi
• 金额: $ 1.79万
• 依托单位: Kanazawa Medical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10670811/
• 关键词: Dermatophytes; Identification; Molecular technique; ribosomal DNA; ITS; リボゾームDNA

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

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）通过Mva I或提示。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。此外，该技术价格低廉，节省时间，因此该方法也适合同时处理多个分离株。总之，我建议使用Mva I或Hinf I对ITS 1+2区域进行PCR-RFLP分析，用于实际中皮肤真菌的鉴定。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

项目成果

Mochizuki,T., Kawasaki,M., Ishizaki,H., Makimura,K.: "The Identification of several clinical isolates of dermatophytes based on the nucleotide sequence if internal transcribed spacer 1)ITS1) in nuclear ribosomal DNA"Journal of Dermatology. Vol. 26. 276-28

Mochizuki,T.、Kawasaki,M.、Ishizaki,H.、Makimura,K.：“根据核核糖体 DNA 中的内转录间隔区 1)ITS1) 的核苷酸序列鉴定皮肤癣菌的几种临床分离株”皮肤病学杂志

Takashi Mochizuki et al.: "Identification of several clinical isolates of dermatophytes based on the nucleotide sequence of internal transcribed spacer 1 in nuclear ribosomal DNA"The Journal of Dermatology. 26. 276-281 (1999)

Takashi Mochizuki 等人：“基于核核糖体 DNA 中内转录间隔区 1 的核苷酸序列鉴定皮肤癣菌的几种临床分离株”《皮肤病学杂志》。

Takashi Mochizuki, et al.: "Identification of several clinical isulates of devoatophytes based on the nucleotidk seguence of Internal transcribed space_r1(ITS 1)in nuclear ribosomal DNA."Journal of Dermatology. 26. 276-281 (1999)

Takashi Mochizuki 等人：“基于核核糖体 DNA 中内部转录空间_r1 (ITS 1) 的核苷酸序列对几种临床分离菌的鉴定。”皮肤病学杂志。

Takashi Mochizuki, et al.: "Identification of several clinical isolates of dermatophytes based on the nucleotide sequence of Internal transcribed spacer 1(ITS 1) in nuclear ribosomal DNA."Journal of Dermatology. 26. 276-281 (1999)

Takashi Mochizuki 等人：“基于核核糖体 DNA 中内部转录间隔区 1 (ITS 1) 的核苷酸序列鉴定皮肤癣菌的几种临床分离株。”皮肤病学杂志。