Menadione-catalyzed luminol chemiluminescent assay for the viability of Escherichia coli ATCC 25922.
Escherichia coli ATCC 25922 produced O2- in the presence of menadione, and O2- -dependent luminol chemiluminescence intensity was proportional to colony-forming unit (CFU) in the exponential phase. CFU was determined by using a 96-well plate at a range of 3 X 10(3) to 8 x 10(7) CFU /well (0.1 ml) after a 10-min incubation with menadione, followed by chemiluminescent assay for 5 s. After a 4-hr incubation of E. coli (10(5) CFU/0.1 ml) with menadione and an antimicrobial agent inhibiting the synthesis of peptidoglycan, protein, and DNA, the inhibitory concentration (IC) of the antimicrobial agent determined by menadione-catalyzed luminol chemiluminescent assay was in good agreement with minimal inhibitory concentration (MIC) of the NCCLS (National Committee for Clinical Laboratory Standard) method requiring 18 hr. Menadione-catalyzed luminol chemiluminescent assay is expected to be useful for the rapid determination of cell viability under the conditions of various cell growths and stresses.
Yamashoji
S
S
Nikken Biomedical Laboratory, Kuze-gun, Kyoto, Japan.
Takeda
M
M
D000583:Amikacin / Q000494:pharmacology; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002384:Catalysis; D015169:Colony Count, Microbial; D004926:Escherichia coli / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D007202:Indicators and Reagents / Q000737:chemistry* / Q000378:metabolism; D008163:Luminescent Measurements; D008165:Luminol / Q000737:chemistry* / Q000378:metabolism; D008826:Microbial Sensitivity Tests / Q000379:methods*; D009249:NADP / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism; D013752:Tetracycline / Q000494:pharmacology; D024483:Vitamin K 3 / Q000737:chemistry*
D000900:Anti-Bacterial Agents; D007202:Indicators and Reagents; D009249:NADP; D008165:Luminol; D024483:Vitamin K 3; D000583:Amikacin; D013752:Tetracycline; D010100:Oxygen