• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Antimicrobial agents and chemotherapy. >Rapid, Semiquantitative Assay To Discriminate among Compounds with Activity against Replicating or Nonreplicating Mycobacterium tuberculosis
【24h】

Rapid, Semiquantitative Assay To Discriminate among Compounds with Activity against Replicating or Nonreplicating Mycobacterium tuberculosis

机译：快速，半定量测定可区分具有抗复制或非复制结核分枝杆菌活性的化合物

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The search for drugs that can kill replicating and nonreplicating Mycobacterium tuberculosis faces practical bottlenecks. Measurement of CFU and discrimination of bacteriostatic from bactericidal activity are costly in compounds, supplies, labor, and time. Testing compounds against M. tuberculosis under conditions that prevent the replication of M. tuberculosis often involves a second phase of the test in which conditions are altered to permit the replication of bacteria that survived the first phase. Falsepositive determinations of activity against nonreplicating M. tuberculosis may arise from carryover of compounds from the nonreplicating stage of the assay that act in the replicating stage. We mitigate these problems by carrying out a 96-well microplate liquid MIC assay and then transferring an aliquot of each well to a second set of plates in which each well contains agar supplemented with activated charcoal. After 7 to 10 days-about 2 weeks sooner than required to count CFU-fluorometry reveals whether M. tuberculosis bacilli in each well have replicated extensively enough to reduce a resazurin dye added for the final hour. This charcoal agar resazurin assay (CARA) distinguishes between bacterial biomasses in any two wells that differ by 2 to 3 log10 CFU. The CARA thus serves as a pretest and semiquantitative surrogate for longer, more laborious, and expensive CFU-based assays, helps distinguish bactericidal from bacteriostatic activity, and identifies compounds that are active under replicating conditions, nonreplicating conditions, or both. Results for 14 antimycobacterial compounds, including tuberculosis (TB) drugs, revealed that PA-824 (pretomanid) and TMC207 (bedaquiline) are largely bacteriostatic.
机译：寻找可以杀死复制型和非复制型结核分枝杆菌的药物面临着实际的瓶颈。 CFU的测量以及从杀菌活性中区分出抑菌剂在化合物，耗材，人工和时间上都是昂贵的。在防止结核分枝杆菌复制的条件下测试化合物对抗结核分枝杆菌的方法通常涉及测试的第二阶段，在该阶段的第二阶段是改变条件以允许在第一阶段幸存的细菌复制。针对非复制性结核分枝杆菌的活性的假阳性测定可能是由于来自测定的非复制阶段的，在复制阶段起作用的化合物的残留。我们通过进行96孔微孔板液体MIC分析，然后将每个孔的等分试样转移到第二组板中来缓解这些问题，在第二组板中，每个孔中都含有琼脂，并添加了活性炭。 7至10天后（比计算CFU荧光法所需的时间早2周），发现每个孔中的结核分枝杆菌是否已充分复制，以减少最后一小时添加的刃天青染料。此木炭琼脂刃天青测定法（CARA）可以区分任何两个相差2至3 log10 CFU的孔中的细菌生物量。因此，CARA可以作为较长，更费力和昂贵的基于CFU的测定的预测试和半定量替代品，有助于区分杀菌活性和抑菌活性，并鉴定在复制条件下，非复制条件下或在这两种条件下均具有活性的化合物。 14种抗分枝杆菌化合物（包括结核病（TB）药物）的结果表明，PA-824（类前驱体）和TMC207（贝达喹啉）在很大程度上具有抑菌作用。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号