Cooksey R C, Morlock G P, Beggs M, Crawford J T
Division of Bacterial and Mycotic Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia, USA.
Antimicrob Agents Chemother. 1995 Mar;39(3):754-6. doi: 10.1128/AAC.39.3.754.
Plasmid pLUC10, carrying the firefly luciferase gene, was transformed by electroporation into Mycobacterium avium A5. Bioluminescence production by strain A5(pLUC10), as measured in a microdilution plate luminometer, was approximately 1 relative light unit per 2 x 10(6) viable bacilli, whereas it was 0.0005 relative light unit for an equal number of parental cells. The susceptibility of strain A5(pLUC10) to eight concentrations of each of eight antimicrobial agents was evaluated by the luciferase microplate assay in parallel with a conventional broth macrodilution method with antimicrobial agents. Decreases in bioluminescence to levels that were < or = 10% of those of drug-free controls were observed in microplate wells containing inhibitory concentrations of drugs in as few as 3 days. The close correlation of these inhibitory concentrations with the MICs determined by a conventional broth macrodilution method suggests that the luciferase microplate method may offer a convenient and reliable means of evaluating the in vitro activities of antimicrobial agents against the M. avium complex.
携带萤火虫荧光素酶基因的质粒pLUC10通过电穿孔法转化到鸟分枝杆菌A5中。在微量稀释板发光计中测量，菌株A5(pLUC10)的生物发光产量约为每2×10(6)个活芽孢杆菌1个相对光单位，而相同数量的亲代细胞为0.0005个相对光单位。通过荧光素酶微孔板分析法，与使用抗菌剂的传统肉汤宏稀释法平行，评估了菌株A5(pLUC10)对八种抗菌剂的八种浓度的敏感性。在含有抑制浓度药物的微孔板孔中，短短3天内就观察到生物发光降低至无药物对照的<或=10%的水平。这些抑制浓度与通过传统肉汤宏稀释法测定的最低抑菌浓度密切相关，这表明荧光素酶微孔板法可能提供一种方便可靠的方法来评估抗菌剂对鸟分枝杆菌复合群的体外活性。
