Optimization of the microplate resazurin assay for screening against Mycobacterium bovis strain BCG 43756

Abstract

Sari M, Arismayanti E, Kusharyoto W. 2016. Optimization of the microplate resazurin assay for screening against Mycobacterium bovis strain BCG 43756. Pros Sem Nas Masy Biodiv Indon 2: 189-192. Microplate resazurin reduction assay (MRRA) is a rapid method to evaluate the natural extract products and the synthetic drug used to suppress anti-Mycobacterium growth using a resazurin dye as an indicator. This study aimed to present a pre-screening optimization of condition that affects mycobacterium growth activity. The detection of MRRA from medicine plant extract using Mycobacterium bovis strain BCG 43756 was conducted for 1-2 days incubation in a microplate with 37oC. Variation assay conditions were evaluated such as resazurin indicator concentration, DMSO solvent concentration and incubation time duration as an anti-mycobacterial activity. The result of pre-screen showed that the optimization of MRRA using 0.5% DMSO solvent and a resazurin dye of 60 μg/mL with an addition of 5% tween 80 for 24 hours incubation was an ideal condition for decreasing mycobacterium activity. The detection of extract using MRRA assay showed a screen model has rapid detection, simple, and good sensitivity for the development of potential natural extracts from Indonesian medicinal plants.