白假丝酵母菌体外生物膜药敏性不同检测方法的比较

Different detection methods for antifungal susceptibility of Saccharomyces albicans biofilms in vitro

目的比较不同的方法对白假丝酵母菌体外生物膜药敏性检测的差异。方法分别采用菌落计数法(CFU)、AlamarBlue试剂法、XTT减低法、MTT法对白假丝酵母菌体外48h成熟生物膜的药敏性进行检测,并将AlamarBlue试剂法、XTT减低法、MTT法与CFU法进行比较,观察其相关性及差异性。结果 AlamarBlue试剂法、XTT减低法、MTT法与CFU法都有较高的相关性,相关系数分别为r=0.969、r=0.971、r=0.982(P<0.01);与CFU法的差异分别为(0.093±0.127)、(0.054±0.113)、(0.013±0.066),其差异无统计学意义(P>0.05)。结论 AlamarBlue试剂法、XTT减低法、MTT法均可替代CFU法对白假丝酵母菌生物膜进行药敏检测。MTT法与CFU法的相关性最大,差异性最小;AlamarBlue试剂作为一种新的试剂,操作简单,对细胞及人类无毒害,更加适合高通量检测。

Objective To compare different detection methods for antifungal susceptibility of Saccharomyces albicans biofilms in vitro.Methods The Colony-Forming Units（CFU）method,AlamarBlue assay,XTT reduction assay and MTT assay were used to detect the antifungal susceptibility of Saccharomyces albicans biofilms（cultured for 48 hour in vitro）respectively;the results were compared in terms of the correlations and differences among the four methods using SPSS system.Results The AlamarBlue assay,XTT reduction assay and MTT assay demonstrated a high correlation with CFU method;the correlation coefficients were r=0.969,r=0.971,and r=982（P0.01）respectively,while the differences were（0.093±0.127）,（0.054±0.113）and（0.013±0.066）（P0.05）respectively.Conclusion AlamarBlue assay,XTT reduction assay and MTT assay can replace the CFU method in the antifungal susceptibility testing of Saccharomyces albicans biofilms.Compared with AlamarBlue assay and XTT reduction assay,the MTT method has higher correlation and smaller difference with CFU method.As a new reagent,AlamarBlue assay is easy to operate and non-toxic to cells and human,and is more suitable for high throughput detection.