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目的体外诱导近平滑假丝酵母菌(CP)、热带假丝酵母菌(CT)对两性霉素B(AMB)的耐药株,检测其中的耐药基因表达情况。方法 AMB体外构建耐药株,用实时荧光定量PCR法检测耐药基因的表达水平。结果 CP、CT野生株经体外AMB浓度递增法诱导后,形成耐药株最低抑菌浓度(MIC,≥8μg·m L-1);CYP51A、CYP51B、CDR1、CDR2、MDR1及MDR2基因在野生株中低水平表达,在耐药株中表达水平明显升高,差别有统计学意义(P<0.05);CYP51A、CYP51B、CDR1、CDR2、MDR1及MDR2的表达量在CPAMB耐药株中分别是其野生株的136.92,16.33,2.07,14.84,4.07,2.38倍;在CT AMB耐药株中分别是其野生株的8.83,2.98,3.00,2.40,2.30,3.57倍。结论 CP、CT对AMB耐药与CYP51A、CYP51B、CDR1、CDR2、MDR1及MDR2基因过度表达有关。
Objective To induce the resistant strains of Candida parapsilosis (CP) and Candida tropicalis (CTB) to amphotericin B (AMB) in vitro and detect the expression of multidrug resistance genes. Methods AMB-resistant strains were constructed in vitro and the expression of drug-resistant genes was detected by real-time fluorescence quantitative PCR. Results The minimum inhibitory concentration (MIC) of CYP51A, CYP51B, CDR1, CDR2, MDR1 and MDR2 gene in wild-type strain of CP and CT strains after being induced by AMB concentration in vitro increased (P <0.05). The expression levels of CYP51A, CYP51B, CDR1, CDR2, MDR1 and MDR2 in CPAMB resistant strains were respectively 136.92,16.33,2.07,14.84,4.07,2.38 times that of the wild-type strain, and 8.83,2.98,3.00,2.40,2.30,3.57 times higher than that of the wild-type CT AMB strain. Conclusions CP and CT are related to the resistance of AMB to overexpression of CYP51A, CYP51B, CDR1, CDR2, MDR1 and MDR2 genes.