Biotechnology and Food Research Institute, Fukuoka Industrial Technology Center, Kurume 839-0861, Japan.
J Microbiol Methods. 2012 Sep;90(3):160-6. doi: 10.1016/j.mimet.2012.05.001. Epub 2012 May 27.
The minimum inhibitory concentrations (MICs) obtained from the susceptibility testing of various bacteria to antibiotics were determined by a colorimetric microbial viability assay based on reduction of a tetrazolium salt {2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt (WST-8)} via 2-methyl-1,4-napthoquinone as an electron mediator and compared with those obtained by the broth microdilution methods approved by the Clinical and Laboratory Standard Institute (CLSI). Especially for drug-resistant bacteria, the CLSI method at an incubation time of 24h tended to give lower MICs. The extension of incubation time was necessary to obtain consistent MICs for drug-resistant bacteria such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococi (VRE) and multi-drug resistant Pseudomonas aeruginosa (MDRP) in the broth microdilution method. There was excellent agreement between the MICs determined after 24h using the WST-8 colorimetric method and those obtained after 48-96 h using the broth microdilution method. The results suggest that the WST-8 colorimetric assay is a useful method for rapid determination of consistent MICs for drug-resistant bacteria.
采用基于 2-甲基-1,4-萘醌作为电子介体还原 2-(2-甲氧基-4-硝基苯基)-3-(4-硝基苯基)-5-(2,4-二磺基苯基)-2H-四唑单钠盐(WST-8)的比色微生物活力测定法,测定了各种细菌对抗生素的药敏试验的最小抑菌浓度(MICs),并与临床和实验室标准协会(CLSI)批准的肉汤微量稀释法获得的 MICs 进行了比较。对于耐药菌,CLSI 法在孵育 24h 时往往会给出较低的 MICs。需要延长孵育时间,才能使肉汤微量稀释法中耐甲氧西林金黄色葡萄球菌(MRSA)、万古霉素耐药肠球菌(VRE)和多药耐药铜绿假单胞菌(MDRP)等耐药菌获得一致的 MICs。采用 WST-8 比色法测定 24h 后的 MICs 与采用肉汤微量稀释法测定 48-96h 后的 MICs 具有极好的一致性。这些结果表明,WST-8 比色法是一种快速测定耐药菌一致 MICs 的有用方法。