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目的比较马尔尼菲蓝状菌(Talaromyces marneffei,T.marneffei)耐药株和敏感株氟康唑(Fluconazole,FCZ)耐药基因在转录水平上表达量的差异,为马尔尼菲青霉病(Penicilliosis marneffei,PSM)临床用药及预后监测提供参考依据。方法采用FCZ浓度递增法对11株酵母相T.marneffei进行体外诱导耐药,按临床实验室标准委员会(NCCLS)颁布的微量稀释法测其最小抑菌浓度(minimal inhibitory concentration,MIC值),通过实时荧光定量逆转录-聚合酶链反应(Real-time fluomgenetic quantitative PCR,FQ-PCR)测定耐药株及敏感株FCZ耐药基因的表达量,比较分析T.marneffei的MIC值变化前后与FCZ耐药基因表达量的关系。结果 11株酵母相T.marneffei经诱导后,8株T.marneffei达到耐药判断标准,FCZ耐药基因在T.marneffei中均表达,表达量由诱导前的2.56±1.84上升至7.50±3.00,差异有统计学意义(P﹤0.05);每株T.marneffei经FCZ诱导后FCZ耐药基因表达量均有不同程度的上升,上升幅度范围在1.8~16.4倍之间。结论采用FCZ浓度递增法可成功在体外诱导酵母相T.marneffei耐药,T.marneffei对FCZ的耐药性可能与FCZ耐药基因的表达有关。
OBJECTIVE: To compare the differences in transcriptional levels of resistance genes of Talaromyces marneffei (T. marneffei) and susceptible fluconazole (FCZ) Penicilliosis marneffei, PSM) clinical medication and prognosis monitoring provide a reference. Methods Eleven strains of yeast T.marneffei were induced in vitro by FCZ method. The minimal inhibitory concentration (MIC) was determined according to the microdilution method promulgated by the National Laboratory Standards Committee (NCCLS) Real-time fluorescence quantitative PCR (FQ-PCR) was used to detect the expression of FCZ resistance in drug-resistant and susceptible strains. The MIC values of T. marneffei were compared with those of FCZ Relationship between drug gene expression. Results After induced by T.marneffei, 8 strains of T.marneffei reached the resistance criteria. The FCZ resistance genes were all expressed in T.marneffei, and the expression increased from 2.56 ± 1.84 before induction to 7.50 ± 3.00, (P <0.05). The expression of FCZ resistance gene of T.marneffei strain was increased by 1.8 ~ 16.4 folds after FCZ induction. CONCLUSION: The yeast-phase yeast T.marneffei is successfully induced by FCZ concentration increasing. The resistance of T. marneffei to FCZ may be related to the expression of FCZ resistance gene.