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目的建立常见病原假丝酵母菌种间分型和对白色假丝酵母菌种内基因分型的方法,并分析白色假丝酵母菌不同基因型与耐药表型的关系。方法采用PCR技术扩增假丝酵母菌r DNA的内转录间隔区ITS1-ITS2,根据PCR产物片段的大小进行假丝酵母菌种间分型;采用特异引物扩增白色假丝酵母菌基因组DNA的25S r DNA基因的内含子区,以内含子大小及其中可转座I型内含子片段的缺失或插入作为分型依据,对白色假丝酵母菌进行种内基因分型。并对肺癌患者感染的白色假丝酵母菌不同基因型的耐药性进行分析。结果依据科玛嘉显色培养基结果和PCR电泳图谱,与基因库中假丝酵母菌基因序列比对,分为四种:白色假丝酵母菌242株大小218 bp,光滑假丝酵母菌101株大小480 bp,热带假丝酵母菌49株大小219 bp,克柔假丝酵母菌16株大小150 bp。临床分离的242株白色假丝酵母菌经PCR方法分为3型:A型166株(450 bp),B型54株(840 bp),C型22株(450、840 bp)。A型为最常见的基因型,B型、C型白色假丝酵母菌对唑类抗真菌药物的耐药率显著高于A型菌株(P<0.05),对其他抗真菌药物的耐药率差异无统计学意义。结论 PCR方法用于假丝酵母菌种间与种内的分型具有较高的灵敏性与特异性,不同型的白色假丝酵母菌耐药性可能与特定基因型有一定相关性。
OBJECTIVE: To establish a method for genotyping Candida albicans and genotyping in Candida albicans, and analyze the relationship between different genotypes of Candida albicans and drug resistance phenotype. Methods The ITS1-ITS2 of Candida albicans r DNA was amplified by PCR. The Candida species was typed according to the size of PCR products. The genomic DNA of Candida albicans was amplified by specific primers Intron region of 25S r DNA gene was genotyped by intraspecific inoculation based on the size of intron and the deletion or insertion of transposable type I intron fragment. The drug resistance of different genotypes of Candida albicans infected with lung cancer patients were analyzed. The results were based on the results of Kema Jia chromogenic medium and PCR electrophoresis and were compared with the Candida albicans gene sequences in the gene bank. There were four types of Candida albicans: 242 strains of Candida albicans, 218 bp in length, Candida glabrata 101 The size of the strain was 480 bp, 49 strains of Candida tropicalis were 219 bp, and 16 strains of Candida krusei were 150 bp in size. A total of 242 clinical isolates of Candida albicans were divided into 3 types by PCR: 166 strains of A (450 bp), 54 strains of B (840 bp) and 22 strains of C (450,840 bp). Type A was the most common genotype. The resistance rate of Candida albicans to azole was significantly higher than that of type A (P <0.05), and the rates of resistance to other antifungal agents The difference was not statistically significant. Conclusion PCR method for Candida species and intraspecific typing with high sensitivity and specificity, different types of Candida albicans drug resistance may be related to certain genotypes.