Bioluminescence screening in vitro (Bio-Siv) assays for high-volume antimycobacterial drug discovery
Bioluminescence screening in vitro (Bio-Siv) assays for high-volume antimycobacterial drug discovery
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DOI：
10.1128/aac.40.6.1536
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发表时间：
1996-06-01
影响因子：
4.9
通讯作者：
Stover, CK
中科院分区：
文献类型：
--
作者：
Arain, TM;Resconi, AE;Stover, CK
已经开发了基于生物发光的检测方法来指示抗菌素敏感性，并对表达真核荧光素酶基因的结核分枝杆菌、牛分枝杆菌、禽分枝杆菌和细胞内分枝杆菌的重组菌株进行了验证。用这些生物发光分析方法测定几种抗分枝杆菌药物，包括异烟肼、乙胺丁醇、利福平、阿米卡星、链霉素、环丙沙星和克拉霉素的最低抑菌浓度，与传统的BACTEC方法和对抑制终点的直观估计相比，分析方法已被优化，用于分析大量新化合物，并且简单、廉价和劳动效率高，这四种重组分枝杆菌的可用性使得使用非致病BCG菌株进行大规模筛选并随后确认对致病分枝杆菌的活性的药物发现策略成为可能。此外，有证据表明，基于BCG的筛选可能允许直接鉴定杀菌剂。
Bioluminescence-based assays to indicate antimicrobial susceptibility have been developed and validated for recombinant strains of Mycobacterium tuberculosis, Mycobacterium bovis BCG, Mycobacterium avium, and Mycobacterium intracellulare expressing an integrated eukaryotic luciferase gene. MICs determined with these bioluminescence assays for several antimycobacterial agents, including isoniazid, ethambutol, rifampin, amikacin, streptomycin, ciprofloxacin, and clarithromycin, compared favorably with traditional BACTEC methods and visual estimations of the inhibitory end point, Assay methodology has been optimized for the analysis of large numbers of novel compounds and is simple, inexpensive, and labor efficient, The availability of these four recombinant mycobacteria has permitted a strategy for drug discovery employing the nonpathogenic BCG strain for mass screening purposes with subsequent confirmation of activity against the pathogenic mycobacteria, Furthermore, evidence suggests that the BCG-based screen may allow the direct identification of bactericidal agents.