【摘要】 目的探讨红色毛癣菌基因型的稳定性。方法采用随机引物PCR法和探针与DNA印迹杂交法。以CTAB法提取DNA。随机引物为OPAA11[5-′ACCCGACCTG-3′],印迹法以NS5和ITS4(真菌的特异性引物)为引物,以红色毛癣菌的标准株为模板,扩增出rDNA的部分18S区、ITSⅠ区、5.8S区和ITSⅡ区为探针,并用随机引物法将探针用P32标记,与EcoRⅠ酶切的基因组DNA杂交。结果①AP-PCR法:红色毛癣菌扩增的主要带型是2.2,1.7,1.3,0.9,0.7 kb,所试红色毛癣菌传代前后扩增带型均无变化。②探针与DNA印迹杂交法:原代(1代)与传代后的(2,3,4代)杂交带型一致,11株红色毛癣菌均表现为3条带,分两型(分子量分别为2.4,3.9,5.9 kb和2.4,4.4,6.5 kb)。结论红色毛癣菌基因型稳定。更多还原

【Abstract】 Objective To explore the stability of genotype in Trichophyton rubrum.Methods DNA was extracted with CTAB method.A probe which consisted of the 3′end of the 18S rDNA,adjacent ITS1,5.8S rDNA and ITS2 regions was amplified from template DNA of the T.rubrum standard strain by using fungal universal primers NS5 and ITS4,labelled by P32 and hybridized with EcoRI-digested T.rubrum genomic DNA.Arbitrarily primed PCR(AP-PCR) was performed using random primer [5′-ACCCGACCTG-3′].Results ①AP-PCR analysis of eleven strains including ten T.rubrum isolates and one T.rubrum standard strain showed similar DNA pattern:2.2,1.7,1.3,0.9,0.7 kb.No changes in DNA pattern were found upon subculture.②From the results of hybridization,three bands and two types(2.4,3.9,5.9 kb and 2.4,4.4,6.5 kb) among the eleven T.rubrum strains were observed.No changes in band pattern was found when the strains were subcultured.Conclusion The genotype of T.rubrum is relatively stable on subculture.更多还原