I used a non-pathogenic fast growing Mycobacteria to screen my natural product compounds for activity. Now I want to test the active compounds in a disk diffusion assay against pathogenic Mycobacterium tuberculosis which grows very slowly, is this still possible to do? Has anyone done this with good results? I am at a university lab that has no funding so I cannot do any anti-Mtb assays that require buying anything, please let me know your suggestions. Thanks!
I suppose you can use the Disk diffusion assay and you might also find several protocols for the same. However, I would recommend you to only perform the test if you have Biosafety level 2 lab.
Since you are using the human pathogen, cultivation, growth and disposal must always be performed under the guidelines.
Virulent MTB is a BSL-3 restricted pathogen...
As for disk diffusion.. yes, you can do it...however...make sure your agar isn't too thin otherwise the plate will dry out. The 7H11 Middlebrook agar plates are usually 15mm thick.
And I agree with above comment, its strictly a BSL-3 organism. Handle only if you have access and training to handle.
Better to put 1mL 7H9 with OADC/ADS with compound at various conc. and cell at a conc. of 1X10^8 cell/ml in tubes. Some standard drug may be used as control (rifampicin, fluoroquinlone). You may visually check afterwards. This may give yiu an idea in 5-7 days, while plate may take 3-4 weeks.
Hope it may help.
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