应用PCR-RFLP进行申克孢子丝菌的分子生物学鉴定

Molecular Biological Identification of Sporothrix Schenckii by PCR-RFLP

目的 探索一种简单、快速的申克孢子丝菌的鉴定方法。方法 应用真菌通用引物ITS1和ITS4对来源于不同地区及不同临床型别孢子丝菌病的 2 8株申克孢子丝菌以及 9种其他临床上重要的真菌进行PCR扩增 ,利用限制性内切酶HaeⅢ对PCR产物进行酶切分析鉴定。结果 所有 2 8株申克孢子丝菌和其他 9种真菌均扩增出一条约 3 5 0bp的片段 ,其中 2 8株申克孢子丝菌RFLP带型一致 ,与 9种其他临床上重要的真菌RFLP带型差异较明显。 结论 PCR RFLP可以为建立一种简单、快速鉴定申克孢子丝菌的方法提供依据。

Objective To explore a simple and rapid method for identification of Sporothrix schenckii. Methods ①The genomic DNA was extracted from 28 strains of Sporothrix schenckii collected from different clinical types and various regions,and 9 other important clinical species including Aspergillus fumigatus,Exophiala jeanselmei,Phialophora verrucosa,Cladosporium carrionii,Cryptococcus neoformans,Candida krusei,Candida albicans,Microsporum gypseum and Penicillium marneffei. ②Amplify genomic DNA of all the strains and species with the universal primers ITS1 and ITS4.③Digest the amplified products with restriction enzyme HaeⅢ.Results ①A 350bp fragment was amplified from all the strains and species with the universal primers ITS1 and ITS4.②An identical PCR RFLP pattern from all the 28 strains of Sporothrix schenckii was different from the other 9 important clinical species.Conclusion The PCR RFLP may serve for a simple and rapid identification of Sporothrix schenckii.