黄曲霉中唑类耐药基因Cyp51突变位点鉴定OA
Identification of the Cyp51 mutation site of the azole resistance gene in Aspergillus flavus
对新疆临床分离的 6 株黄曲霉进行分子鉴定及唑类耐药基因cyp51A、cyp51B和cyp51C耐药突变表型鉴定.采用PCR扩增ITS和BenA基因,扩增产物进行测序,blast比对后进行菌种的分子鉴定;采用Sensititre Yeastone真菌药敏试剂盒检测7种药物的体外抗真菌药物敏感性,MIC/MEC结果解释标准依据 CLSI M57S 推荐;设计特异性引物,采用 PCR 扩增黄曲霉菌株的 cyp51A、cyp51B和 cyp51C基因并测序,测序结果与黄曲霉参考菌株比对鉴定耐药突变表型.真菌药敏试验MIC/MEC值显示,6 株菌株均对棘白菌素类高度敏感(MEC≤0.015 μg/mL)、对伊曲康唑和泊沙康唑敏感率为100%;1株对两性霉素B耐药(MIC=8 μg/mL),2株对伏立康唑耐药(MIC=4 μg/mL).基因测序结果显示,cyp51A和cyp51C在敏感株和耐药株中均出现了同义和非同义点突变,cyp51B中未发现突变.然而,cyp51C 基因中 P276T 突变位点只存在于耐药菌株中.棘白菌素类、伊曲康唑和泊沙康唑对黄曲霉的体外抑菌活性较好,伏立康唑和两性霉素对黄曲霉的抑菌活性较差;cyp51C P276T突变可能是黄曲霉对伏立康唑耐药的新潜在机制.
The molecular characterization and phenotype identification of resistance mutations of azole resistance genes cyp51A,cyp51B and cyp51C of 6 clinical Aspergillus flavus isolates from Xinjiang are carried out.The isolates were identified based on internal transcribed spacer(ITS)and BenA gene sequencing.The Sensititre Yeastone Fungal Drug Sensitivity Kit was used to detect the in vitro antifungal susceptibility of 7 drugs,and the MIC/MEC results were interpreted according to the standards recommended by CLSI M57S.The cyp51A,cyp51B and cyp51C genes of A.flavus strains were amplified and sequenced by PCR,and the sequencing results were compared with the reference strain of A.flavus to identify the resistance mutation phenotype.All 6 clinical isolates were identified as A.flavus.The results of antifungal susceptibility test showed that the isolated A.flavus strains were completely sensitive to echinocandins,itraconazole and posaconazole;of which 1 strain was resistant to amphotericin(8 μg/mL)and 2 strains were resistant to voriconazole(4 μg/mL).Sequencing of the cyp51A,cyp51B,and cyp51C genes of 2 azole-sensitive and 4 azole-resistant strains revealed that both cyp51A and cyp51C in sensitive and resistant strains showed synonymous and non-synonymous point mutations,and no mutation was found in cyp51B.However,the P276T mutation site in cyp51C gene was only found in one resistant isolate.Echinocandins,itraconazole and posaconazole showed better antifungal activity against A.flavus,while voriconazole and amphotericin showed lower antifungal activity against A.flavus.Probably cyp51C gene mutation of A.flavus is associated with azole resistance.
艾柯代·玉苏甫;祖拜旦木·艾则孜;哈地利亚·哈斯木;王晓东
新疆医科大学第一附属医院皮肤科,新疆 乌鲁木齐 830011新疆医科大学第一附属医院皮肤科,新疆 乌鲁木齐 830011新疆医科大学第一附属医院皮肤科,新疆 乌鲁木齐 830011新疆医科大学第一附属医院皮肤科,新疆 乌鲁木齐 830011
黄曲霉抗真菌药物敏感性唑类耐药cyp51基因
Aspergillus flavusantifungal susceptibilityazole-resistancecyp51 gene
《菌物学报》 2026 (2)
110-116,7
新疆维吾尔自治区"天山英才医药卫生高层次人才培养计划中青年骨干人才"(TSYC202301B029) This work was supported by the Xinjiang Uygur Autonomous Region"Tianshan Elite Medical and Health High-level Talent Training Program Young Backbone Talents"(TSYC202301B029).
评论