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目的探讨热带念珠菌靶位酶(ERG11)基因的改变与氟康唑耐药的相关性。方法根据热带念珠菌ERG11基因序列设计引物,以标准株为对照,对临床分离的耐药株、剂量依赖株(S-DD)和敏感株进行靶位酶ERG11基因PCR扩增,测序之后比对分析标准株,耐药株、剂量依赖株和敏感株的序列差异,分析靶位酶基因改变与氟康唑耐药的相关性。结果热带假丝酵母氟康唑敏感株与敏感标准株序列一致;而S-DD株中出现T783C和G1362A沉默突变;耐药株中出现碱基序列T225C、C461T、G1362A等3种突变点,并导致丝氨酸(S)154苯丙氨酸(F)氨基酸突变。结论热带念珠菌靶位酶ERG11基因氨基酸序列的突变与氟康唑耐药有一定的相关性。
Objective To investigate the relationship between the alteration of ERG11 gene and fluconazole resistance in Candida tropicalis. Methods According to the sequence of ERG11 gene of Candida tropicalis, primers were designed and the standard strain was used as a control. The ERG11 gene was amplified by PCR from the clinically isolated drug-resistant strains, dose-dependent strains (S-DD) and sensitive strains. The sequence differences between standard strains, drug-resistant strains, dose-dependent strains and susceptible strains were analyzed, and the correlation between the target gene changes and fluconazole resistance was analyzed. Results Fluconazole-sensitive strains of Candida tropicalis were consistent with those of sensitive standard strains. However, T783C and G1362A silencing mutations were found in S-DD strains. Three kinds of mutations were found in resistant strains, including T225C, C461T and G1362A Resulting in a mutated amino acid of serine (S) 154 phenylalanine (F). Conclusion The mutation of the amino acid sequence of ERG11 gene in Candida tropicalis has a certain relationship with fluconazole resistance.