Most studies utilise the checkerboard technique or time kill assays to investigate synergistic relationships but neither of these are particularly high throughput. Can anyone suggest a method for testing hundreds of different combinations that is both quick and reliable. My initial idea is to you a modified disk diffusion assay (one antimicrobial in the culture media, the other on a disc) and then following up the best hits with a checkerboard / time kill assay , however my feeling is as these techniques are so different the results may not correlate well. Are there any other techniques that people use?
You can use the Epsilometer or E test assy for your experiment. U can get a details about E test here..... http://aac.asm.org/content/40/8/1914.short
Hi Martin,
I would support your method, add one compound in agar media and another in disk (or well). This would correlate well with checkerboard, simply because both work on the killing or inhibition of the bacteria (provided your drugs do not have diffusibility issues in agar). However, this method may or may not correlate with time-kill assay as it is based on bactericidal property of the drug. So a bacteriostatic drug would give good zones (diffusion assay) and synergy (checkerboard) but fail to show killing in time-kill experiment. Nevertheless, the diffusion assay would be a good primary screen. All the best.
you can perform synergistic activity MIC and MBC by broth dilution method in 96 well titer plate method
After MIC measurement best combination result further analyse by Time kill assay at 1* MIC or 2 * MIC concentration combination.
For detail check this paper
You could consider a luminogenic high throughput system, that utilizes ATP of living organisms in order to produce oxyluciferin from Luciferin. The amount of light produced would then correlate with your living microbes. There are tests out there that use this system to analyze the microbial contamination of food.
So why not put this into a 96 well format?
Have a look at this:
http://multimedia.3m.com/mws/media/686753O/clean-trace-atp-rlus-and-cfus.pdf
and this:
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2846007/
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