An improved MTT colorimetric method for rapid viable bacteria counting

Simple item page
dc.citation.volume214
dc.contributor.authorXu W
dc.contributor.authorShi D
dc.contributor.authorChen K
dc.contributor.authorPalmer J
dc.contributor.authorPopovich DG
dc.coverage.spatialNetherlands
dc.date.accessioned2024-09-08T21:28:25Z
dc.date.available2024-09-08T21:28:25Z
dc.date.issued2023-11
dc.description.abstractThe 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay has been employed in the analysis of bacterial growth. In comparison to experiments conducted on mammalian cells, the MTT bacterial assay encounters a greater number of interfering factors and obstacles that impact the accuracy of results. In this study, we have elucidated an improved MTT assay protocol and put forth an equation that establishes a correlation between colony-forming units (CFU) and the amount of formazan converted by the bacteria, drawing upon the fundamental principle of the MTT assay. This equation is represented as CFU=kF. Furthermore, we have explicated a methodology to determine the scale factor "k" by employing S. aureus and E. coli as illustrative examples. The findings indicate that S. aureus and E. coli reduce MTT by a cyclic process, from which the optimal reduction time at room temperature was determined to be approximately 30 mins. Furthermore, individual E. coli exhibits an MTT reduction capacity approximately four times greater than that of S. aureus. HPLC analysis proves to be the most accurate method for mitigating interferences during the dissolution and quantification of formazan. Additionally, this study has identified a new constraint related to the narrow linear range (0-125 μg/mL) of formazan concentration-absorbance and has presented strategies to circumvent this limitation.
dc.description.confidentialfalse
dc.edition.editionNovember 2023
dc.format.pagination106830-
dc.identifier.author-urlhttps://www.ncbi.nlm.nih.gov/pubmed/37805093
dc.identifier.citationXu W, Shi D, Chen K, Palmer J, Popovich DG. (2023). An improved MTT colorimetric method for rapid viable bacteria counting.. J Microbiol Methods. 214. (pp. 106830-).
dc.identifier.doi10.1016/j.mimet.2023.106830
dc.identifier.eissn1872-8359
dc.identifier.elements-typejournal-article
dc.identifier.issn0167-7012
dc.identifier.number106830
dc.identifier.piiS0167-7012(23)00164-1
dc.identifier.urihttps://mro.massey.ac.nz/handle/10179/71427
dc.languageeng
dc.publisherElsevier BV
dc.publisher.urihttps://www.sciencedirect.com/science/article/pii/S0167701223001641
dc.relation.isPartOfJ Microbiol Methods
dc.rights(c) The author/sen
dc.rights.licenseCC BY-NC-NDen
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/en
dc.subjectEscherichia coli
dc.subjectFormazan
dc.subjectHPLC
dc.subjectMTT bacterial assay
dc.subjectStaphylococcus aureus
dc.subjectViable count
dc.subjectAnimals
dc.subjectColorimetry
dc.subjectFormazans
dc.subjectEscherichia coli
dc.subjectStaphylococcus aureus
dc.subjectTetrazolium Salts
dc.subjectMammals
dc.titleAn improved MTT colorimetric method for rapid viable bacteria counting
dc.typeJournal article
pubs.elements-id480832
pubs.organisational-groupOther
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