白色假丝酵母菌的随机扩增多态性DNA分型研究

Randomly Amplified Polymorphic DNA Study of Candida albicans

目的建立医院感染白色假丝酵母菌的基因多态性DNA分型方法,了解白色假丝酵母菌医院感染的分子流行病学情况,并探讨其基因多态性与真菌药物敏感性检测结果的可能关系。方法收集临床各病区送检的住院患者白色假丝酵母菌感染标本30份和其相应患者资料、抗真菌药物敏感性检测结果,采用经优化的随机扩增多态性(RAPD)DNA方法,对30株白假丝酵母菌进行基因多态性分型,利用Phylip3.67聚类分析软件分别用邻接法和算术平均数的非加权成组配对法,对RAPD扩增条带结果进行聚类分析。结果随机引物OPE-03进行RAPD扩增的指纹图谱清晰,带型稳定,多态性丰富,可以作为分型引物;30株不同来源的菌株可大致分为3种亲缘关系;两种聚类分析结果相同,提示该实验结果可靠。结论白色假丝酵母菌基因多态性分布可能与菌株的来源有关;RAPD多态性可能与其耐药性有关;不同聚类分析方法的RAPD图谱分析结果相同。

OBJECTIVE To genotype the clinical Candida albicans isolates by randomly amplified polymorphic DNA （RAPD） method, study molecular epidemiology of C. albicans infection and discuss possible relationship between RAPD patterns and antifungal test results. METHODS Thirty C. albicans isolates with corresponding patients information and antifungal drug susceptibility test results were collected. All the isolates were genotyped by RAPD fingerprinting by employing primer OPE-03. Cluster analysis was performed by Neighbor Joining and unweighted pair-group method using anarithmetic average methods. RESULTS Primer OPE-03 was suitable for fingerprinting analysis, yielding well-resolved banding patterns. Generally, the isolates could be divided into three clusters. CONCLUSIONS RAPD fingerprinting can be used to genotype C. albicans. The RAPD patterns of C. albicans may be related to the origins of fungi, which also related to drug resistance.