CAJ | 학술논문

目的明确白念珠菌ERG3基因突变及高表达对抗真菌药物耐药的调控作用.方法用微量稀释法测得36株临床分离白念珠菌的最小抑菌浓度(MIC);提取基因组DNA,PCR基因扩增,将扩增后的产物纯化测序,并与genbank中已知的标准序列(AF069572)进行BLAST比对分析;采用实时定量PCR(RT-PCR)方法,对白念珠菌ERG3基因表达的mRNA进行相对定量分析.结果 36株白念珠菌ERG3基因序列共发现6个突变位点,其中19株发生同义突变,突变位点为T51C、T432C、T381C、C438T、T1044C;2株发生错义突变,突变位点为C1052T,并产生耐药;白念珠菌对氟康唑耐药组ERG3高表达的发生率高于敏感组(P＜0.05).结论 ERG3基因突变及高表达增加了抗真菌药物的耐药性.同时有多种调节机制共同参与白念珠菌耐药性的形成.
목적 명학백념주균ERG3기인돌변급고표체대항진균약물내약적조공작용.방법 용미량희석법측득36주림상분리백념주균적최소억균농도(MIC);제취기인조DNA,PCR기인확증,장확증후적산물순화측서,병여genbank중이지적표준서렬(AF069572)진행BLAST비대분석;채용실시정량PCR(RT-PCR)방법,대백념주균ERG3기인표체적mRNA진행상대정량분석.결과 36주백념주균ERG3기인서렬공발현6개돌변위점,기중19주발생동의돌변,돌변위점위T51C、T432C、T381C、C438T、T1044C;2주발생착의돌변,돌변위점위C1052T,병산생내약;백념주균대불강서내약조ERG3고표체적발생솔고우민감조(P＜0.05).결론 ERG3기인돌변급고표체증가료항진균약물적내약성.동시유다충조절궤제공동삼여백념주균내약성적형성.
Objective To investigate the correlation between the mutation and high expression of ERG3 to antifungal drug resistance in candida albicans.Methods Candida albicans was measured minimum inhibitory concentration (MIC) by microdilution susceptibility testing.DNA was extracted.PCR was carried out to amply the full-length ERG3.The purified ERG3 compared with known standard sequences in genbank (AF069572) through BLAST analysis.The mRNA expression of ERG3 was determined by reverse transcription-polymerase chain reaction (RT-PCR).Results Six mutations were detected in ERG3 gene of thirty-Six candida albicans strains.Nineteen strains appeared synonymous-mutations,including T51C,T432C,T381C,C438T,T1044C.Two strains appeared missense mutation,including C1052T.High expression rate of fluconazole resistant group was higher than that of the sensitive group.Conclusion The mutation and high expression of ERG3 genes increased antifungal drug resistance.Meanwhile,variety of regulating mechanism participate in the formation of drug resistance in candida albicans.