Please use this identifier to cite or link to this item: http://hdl.handle.net/1959.14/17107
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- Title
- Optimisation of the fluorescein diacetate antibacterial assay
- Related
- Journal of microbiological methods, Vol. 60 , Issue 1, p.21-30
- DOI
- 10.1016/j.mimet.2004.08.010
- Publisher
- Elsevier
- Date
- 2005
- Author/Creator
- Wanandy, Stephanus
- Author/Creator
- Brouwer, Nynke
- Author/Creator
- Liu, Qian
- Author/Creator
- Mahon, Andrew
- Author/Creator
- Cork, Stephen
- Author/Creator
- Karuso, Peter
- Author/Creator
- Vemulpad, Subramanyam
- Author/Creator
- Jamie, Joanne
- Description
- The fluorescein diacetate (FDA) antibacterial assay relies on the cleavage of fluorescein diacetate by metabolically active bacteria. The recent finding that microbiological media can lead to significant levels of cleavage has reduced the reliability of the assay. Using the nucleophilic scavengers N-ethylmaleimide and maleic anhydride, we have demonstrated that this abiotic cleavage is most likely due to nucleophiles such as cysteine and histidine commonly present in the media. To increase the reliability of the assay we have modified the original assay conditions to include use of dilute medium (peptone 0.2% w/v, yeast extract 0.1% w/v and NaCl 0.1% w/v) in a non-nucleophilic buffer and overnight incubation of the medium after addition of antibacterial agents. The optimised fluorescein diacetate assay has been used to determine the MIC of gentamicin, tetracycline and chloramphenicol for Escherichia coli, Staphyloccocus aureus and Pseudomonas aeruginosa and gave quantitative results that were reproducible and consistent with published data.
- Description
- 10 page(s)
- Subject Keyword
- antibacterial assay
- Subject Keyword
- fluorescein diacetate
- Subject Keyword
- abiotic cleavage of fluorescein diacetate
- Resource Type
- journal article
- Organisation
- Macquarie University. Dept. of Chemistry and Biomolecular Sciences
- Organisation
- Macquarie University. Dept. of Health and Chiropractic
- Organisation
- Macquarie University. Dept. of Biological Sciences
- Identifier
- http://hdl.handle.net/1959.14/17107
- Identifier
- ISSN:1872-8359
- Identifier
- mq-rm-2005000845
- Language
- eng
- Reviewed
