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3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法是一种用于检测查耳酮体外抗结核活性的快速、廉价的筛选试验。

The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay is a rapid, cheap, screening test for the in vitro anti-tuberculous activity of chalcones.

作者信息

Moodley Suventha, Koorbanally Neil A, Moodley Thrineshen, Ramjugernath Deresh, Pillay Manormoney

机构信息

Medical Microbiology and Infection Control, School of Laboratory Medicine and Medical Science, University of KwaZulu-Natal, Durban, South Africa.

School of Chemistry and Physics, University of KwaZulu-Natal, Private Bag X54001, Durban, 4000, South Africa.

出版信息

J Microbiol Methods. 2014 Sep;104:72-8. doi: 10.1016/j.mimet.2014.06.014. Epub 2014 Jun 28.

Abstract

Rapid and reliable drug susceptibility testing facilitates replenishment of the TB drug pipeline in the fight against drug resistant Mycobacterium tuberculosis. This study compared the performance of the MTT and MABA assays on the anti-tuberculous activity of a set of chalcones. Twenty seven chalcones and chromenochalcones were screened against the laboratory strain M. tuberculosis H37Rv, using a microtitre plate MTT assay at 7 days. The MIC for 20 active compounds was subsequently determined using the MABA, MTT and the Macroscopic broth assays at 7, 14 and 21 days. No significant difference in the MICs, or increase in the MICs was observed over time between the MABA (p=0.209) and the MTT (p=0.207) assays, in contrast to the gold standard, the Macroscopic broth assay (p=0.000). The MICs (16 to >128μg/ml) were much higher than the currently used TB drugs. In conclusion, the MTT assay is a cost effective method (R0.06/well) for the rapid in vitro screening of chalcones against M. tuberculosis, producing reliable results in 8 days. The chalcone with a MIC of 16μg/mL shows promise as a potential lead compound and should be investigated further.

摘要

快速可靠的药敏试验有助于在抗击耐药结核分枝杆菌的斗争中补充结核病药物储备。本研究比较了MTT和MABA试验对一组查耳酮抗结核活性的表现。使用微量滴定板MTT试验在7天时针对实验室菌株结核分枝杆菌H37Rv筛选了27种查耳酮和色烯查耳酮。随后在7、14和21天时使用MABA、MTT和宏观肉汤试验测定了20种活性化合物的最低抑菌浓度(MIC)。与金标准宏观肉汤试验(p=0.000)相比,MABA试验(p=0.209)和MTT试验(p=0.207)之间在MIC方面未观察到显著差异,也未观察到MIC随时间增加。MIC(16至>128μg/ml)远高于目前使用的结核病药物。总之,MTT试验是一种经济有效的方法(每孔0.06兰特),用于快速体外筛选查耳酮对结核分枝杆菌的活性,在8天内产生可靠结果。MIC为16μg/mL的查耳酮显示出作为潜在先导化合物的前景,应进一步研究。

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