71株白念珠菌临床分离株微卫星基因分型

Rapid microsatellite genotyping of 71 clinical Candida albicans isolates

目的建立高效、快速、简便的白念珠菌临床分离株三重保守基因微卫星分型技术。方法以煮沸法处理的71株白念珠菌野生株为模板,三色荧光分别标记保守基因CDC3、EF3和HIS3微卫星序列引物,PCR扩增,产物行聚丙烯酰胺凝胶电泳。由基因扫描分析软件获取片段的精确长度,Genotyper软件行基因分型。结果71株白念珠菌野生株行CDC3、EF3和HIS3基因分型分别获得5、10、8种等位基因,7、8、9种基因型,三位点联合分析共获得14种菌株基因型。结论白念珠菌多重保守基因微卫星分型能从基因水平快速、高效的对临床分离株进行分型,对白念珠菌的流行病学研究有重要价值。

Objective To establish efficient, rapid and convenient three microsatellite markers genotyping methord of clinical Candida albicans （C. albicans） isolates. Methods With 71 boiled clinical C. albicans isolates as polymerase chain reaction （PCR） templates, microsatellite sequences of three stable gene loci CDC3, EF3 and HIS3 were respectively amplified by three sets of primers 5＇labeled with different fluoresceins. PCR products were run on acrylamide urea gel and exact sizes of products were read with GeneScan software followed by genotyping with Genotyper software. Results By microsatellite genotyping for 71 clinical C. albicans isolates, S alleles and 7 alleles associations in CDC3 locus, 10 alleles and 8 allele associations in EF3 locus, 8 alleles and 9 allele associations in HIS3 locus were got. Fourteen allele associations were attained by analyzing three loci simultaneous- ly. Conclusions Three microsatellite markers genotyping methord for C. albicans, which can genotype the isolates quickly and accurately in genetic level, has significances in epidemiologic researches.