PCR-RFLP鉴别临床常见的皮肤癣菌

Species Identification of Common Dermatophyte Fungi by PCR-RFLP Analysis of Ribosomal-DNA Intergenic Spacer Regions

目的 建立一种快速而简便的分子水平上鉴定常见皮肤癣菌的方法。方法 采用通用引物ITS1(5’ TCCGTAGGTGAACCTGCGG 3’)、ITS4(5’ TCCTCCGCTTATTGATATGC 3’)对 7种常见皮肤癣菌的核糖体的ITS(in ternaltranscribedspacer)区 (包含 5 .8SrDNA)进行PCR扩增 ,PCR产物分别应用限制性内切酶MvaI、TaqI、HinfI酶切。 结果 7种皮肤癣菌的ITS区扩增产物条带大小不同 ;以MvaI、TaqI、HinfI三种酶分别酶切PCR ITS区产物 ,7种皮肤癣菌均产生独特且易于区分的酶切图 ,尤其是MvaI酶产生的酶切图在种间差异最为明显。可将 7种菌明确区分开。结论 核糖体ITS区PCR RFLP分析是区分常见皮肤癣菌的有价值的方法。

Objective To develop a rapid and simple method for molecular species identification of common dermatophyte fungi.Methods The contiguous ITS and 5.8SrDNA regions were amplified from common dermatophyte species by using the universal primers ITS1(5' TCCGTAGGTGAACCTGCGG 3'),ITS4(5' TCCTCCGCTTATTGATATGC 3').The amplified ITS products were digested with the restriction endonuclease MvaI,TaqI and HinfI.Results The amplified ITS products obtained from 7 dermatophyte species were of different size;Digestion of the amplified ITS products with the restriction endonuclease Mval,TaqI and HinfI produced unique and easily identifiable fragment patterns for 7 dermatophyte species,especially the MvaI restriction endonuclease gave more distinct fragment patterns among these species;7 dermatophyte species could be identified definitaly.Conclusion We conclude that PCR-RFLP analysis of ribosomal-DNA intergenic spacer regions is a valuable method for species identification of common dermatophyte fungi.