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利用随机扩增多态性DNA和rDNA非转录间隔区分析对红色毛癣菌进行菌株鉴别

Strain differentiation of Trichophyton rubrum by randomly amplified polymorphic DNA and analysis of rDNA nontranscribed spacer.

作者信息

Baeza Lilian Cristiane, Matsumoto Marcelo Teruyuki, Almeida Ana Marisa Fusco, Mendes-Giannini Maria José Soares

机构信息

Departamento de Análises Clínicas, Faculdade de Ciêlncias Farmacêuticas, UNESP, Rua Expedicionários do Brasil, 1621, Araraquara, SP, CEP 14801-902, Brazil.

出版信息

J Med Microbiol. 2006 Apr;55(Pt 4):429-436. doi: 10.1099/jmm.0.46236-0.

Abstract

Trichophyton rubrum is the most common pathogen causing dermatophytosis. Molecular strain-typing methods have recently been developed to tackle epidemiological questions and the problem of relapse following treatment. A total of 67 strains of T. rubrum were screened for genetic variation by randomly amplified polymorphic DNA (RAPD) analysis, with two primers, 5'-d[GGTGCGGGAA]-3' and 5'-d[CCCGTCAGCA]-3', as well as by subrepeat element analysis of the nontranscribed spacer of rDNA, using the repetitive subelements TRS-1 and TRS-2. A total of 12 individual patterns were recognized with the first primer and 11 with the second. Phylogenetic analysis of the RAPD products showed a high degree of similarity (>90 %) among the epidemiologically related clinical isolates, while the other strains possessed 60 % similarity. Specific amplification of TRS-1 produced three strain-characteristic banding patterns (PCR types); simple patterns representing one copy of TRS-1 and two copies of TRS-2 accounted for around 85 % of all isolates. It is concluded that molecular analysis has important implications for epidemiological studies, and RAPD analysis is especially suitable for molecular typing in T. rubrum.

摘要

红色毛癣菌是引起皮肤癣菌病最常见的病原体。最近已开发出分子菌株分型方法来解决流行病学问题以及治疗后复发的问题。通过随机扩增多态性DNA(RAPD)分析,使用两种引物5'-d[GGTGCGGGAA]-3'和5'-d[CCCGTCAGCA]-3',以及通过对rDNA非转录间隔区的亚重复元件分析,使用重复亚元件TRS-1和TRS-2,对总共67株红色毛癣菌进行了遗传变异筛选。用第一种引物共识别出12种个体模式,用第二种引物识别出11种。对RAPD产物的系统发育分析表明,在流行病学相关的临床分离株中相似度很高(>90%),而其他菌株的相似度为60%。TRS-1的特异性扩增产生了三种菌株特征性条带模式(PCR类型);代表一份TRS-1和两份TRS-2的简单模式占所有分离株的约85%。结论是分子分析对流行病学研究具有重要意义,RAPD分析特别适合于红色毛癣菌的分子分型。

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