I’d like to know how to prepare the standards strains broth in the best way and how to do the test and interpreter the results
I know that it should be done in triplicates but I’m not sure about the best way of doing the experiment
Any paper suggestions please
Thanks in advance
I would recommend microdilution method using rapid INT colorimetric assay. You can check the literature from this link https://doi.org/10.1016/j.jpha.2015.11.005
It is depends from the nature of bacteria and components.
If the components are characterized by good solubility in water buffers and bacteria are aerobic, you can easly use the standard broth microdilution test, similarly what is reported for common antibiotics in the attached paper.
CLSI MIC determination is a standardized method. A Copy of their 2017 is attached
MIC method https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4023284/
Manuel's explanation is right. If you can use a microdilution test in your experiments, could see this paper:
DOI: http://dx.doi.org/10.15381/rpb.v23i3.12866
Best Regards
http://dx.doi.org/10.15381/rpb.v23i3.12866
Article Inhibición bacteriana por cobre nanoestructurado Actividad i...
Hi Mohamed,
I agree with checking the CLSI protocols of microdilution methods. Also, you can slightly modify the protocol by using resazurin, a blue dye which turns bright pink and then transparent when reduced by bacterial activity. I recommend you to use the standard microdilution method from CLSI adding, after incubation and turbidity measurment, 10 uL of resazurin solution at 0.01% (filter sterilized water solution of resazurin). Then, re-incubate the plate for one hour and then read the MIC. In this way MIC is defined as the minimal concentration of your extract in which there was not color change from blue to pink or transparent.
Also, the pink dye resulting from resazurin reduction is fluorescent, so you can use it for quantification.
Best regards!
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4853446
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1895922/
If you want more details on Alamar Blue modified CLSI-M 27A please let us know.
Consult CLSI Guidelines and may also see. https://www.slideshare.net/singh_br1762/methods-for-determining-fractional-inhibitory-concentration-fic
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