用甲基噻唑蓝筛选抗铜绿假单胞菌药物方法的建立

A new method of screening candidate drugs against Pseudomonas aeruginosa by MTT

目的:建立适合铜绿假单胞菌的定量、快速、体外高通量药物筛选方法。方法:优化甲基噻唑蓝法检测活细菌数量的实验条件,用MIC方法与MTT法比较2种已知抗生素的抗菌作用。结果:微量培养对细菌生长无影响,最佳MTT浓度和反应时间分别为5 mg·mL-1和1 h。在同一培养时间,菌浓度在1×107～1×1010CFU·mL-1范围内,甲臜的吸收度与菌浓度呈线性相关。用2种抗生素比较MIC方法与MTT法药敏测定的结果一致。结论:MTT微量法进行抗铜绿假单胞菌新药筛选操作简便,省时省力,成本较低,结果稳定,具有可重复性,适宜体外高通量药物筛选。

Objective： To establish a high-throughput model for screening candidate agents against Pseudomonas aeruginosa in vitro. Methods： A MTT method was studied to optimize detection of antimicrobial activities against Pseudomonas aeruginosa. The antimicrobial activities of two known antibiotics were compared by the conventional assay and the MTT assay. Results： The micropropagation showed no effects on the bacterial growth. The optimal detection condition was to incubate the cultures with MTT at the concentration of 5 mg·mL^-1 for 1 hour. In the same growth phases, the linearity of living bacterial biomass with formazan＇s absorbance was found within 1 × 10^7 ～1 × 10^10CFU·mL^-1. In the test for antimicrobial activities of the two known antibiotics, the MTT assay and the MIC assay showed the same results. Conclusion： The cost-effective, reliable and reproducible MTT microanalysis may be used for high-throughput screening of drugs.