PCR指纹图技术鉴定临床常见酵母菌的探讨

Identification of Common Yeast Species by DNA Fingerprinting with PCR

目的 探讨PCR指纹图技术用于鉴定临床常见酵母菌的方法。方法 采用PCR方法 ,使用随机引物M 13和T3B对临床常见的 7属 17种酵母菌进行扩增。结果 17种酵母菌的扩增产物在数量和大小上都有其自身特征 ;不同种间酵母菌其PCR指纹图型存在明显的差异 ;对同种酵母菌内不同菌株扩增的结果表明 ,酵母菌的PCR指纹图分析种间变异远远大于种内变异 ,据此能将各种酵母菌有效地区分开。结论 控制好实验条件 ,PCR指纹图技术可有效地鉴定临床常见的酵母菌。

OBJECTIVE To study the method of identification of common yeast species by DNA fingerprinting with PCR. METHODS DNA fingerprinting in different species and strains of medically important yeasts were analyzed by PCR with arbitrary primer: the M13 phage core sequence (5'-GAG GGT GGC GGT TCT-3') and the T3B inter-genic spacer repeat of tRNA (5'-AGG TCG CGG GTT CGA ATC C-3'). The DNA from 7 genera of yeasts including 17 species was amplified by PCR with two primers. RESULTS Distinctive and reproducible sets of amplification products were observed for 11 different Candida and 6 other fungal species. The numbers and sizes of the amplification products were characteristic for each species. All yeast species tested could be clearly distinguished by their amplification patterns. With all primers, PCR fingerprints displayed intraspecies variability. However, PCR profiles obtained from different strains of the same species were far more similar than those derived from different yeasts species. CONCLUSIONS DNA fingerprinting with PCR on suitable conditions can identify the clinical common yeasts.