摘要目的 评价多个位点的PCR特异扩增法和限制性片段长度多态性(RFLP)分析在鉴定新型隐球菌AD型及其杂合类型中的作用.方法 采用针对交配型位点内STE20、MF基因及交配型位点外CLA4、GPal基因的PCR特异扩增法和针对交配型位点外g6341基因的RFLP分析对不同杂合类型的AD型菌株进行鉴定及杂合类型的分析.结果 针对STE20基因的特异扩增法能准确鉴定所有AD型菌株的交配类型,但无法鉴定H型菌株.针对CL44基因的特异扩增法在鉴定AD型菌株上优于GPal基因,针对g6341基因的RFLP分析根据A位点交配型的不同将AD型菌株分为2种RFLP谱型.CLA4、GPal和g6341基因不能用于鉴定AD型的交配类型,且可能存在A型或D型位点的缺失.结论 新型隐球菌AD型菌株及其杂合类型的准确鉴定须结合针对交配型位点内外基因的多位点分析.

abstractsObjective To identify the AD hybrid strains and its hybrid types within Cryptococcus neoformans.Methods Difierent hybrid types of AD strains were analyzed by PCR 0f STE20 and MF genes within MAT locus and CIA4 and GPal genes out of MAT locus.The PCR-RFLP analysis of g6341 gene was also performed.Results The mating types of 18 AD strains were precisely identified by PCR of STE20 gene,whereas those of H strain were not identified.CL44 gene was better than the GPal gene in PCR identification of the AD hybrids.In the RFLP analysis of g6341 gene,AD strains were grouped into 2 distinct RFLP patterns based on the mating type on serotype A allele.The mating types of AD strains were not identified by the molecular analyses based on the CL44,GPal and g6341 genes.Conclusion It is necessary to use multi-locus analyses of genes within and out of the MAT locus in precise identification of the AD strains and their hybrid types of Cryptococcus neoformans.