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BioAssay: AID 651850

Inhibition of Mycobacterium tuberculosis during logarithmic growth, as enumerated by colony forming units_2157-05_Inhibitor_Dose_DryPowder_Activity

This assay determines the inhibitory effect of small molecules against M. tuberculosis under replicating conditions ..more
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 Tested Compounds
 Tested Compounds
All(27)
 
 
Active(19)
 
 
Inactive(9)
 
 
 Tested Substances
 Tested Substances
All(28)
 
 
Active(19)
 
 
Inactive(9)
 
 
 Related BioAssays
 Related BioAssays
AID: 651850
Data Source: Broad Institute (2157-05_Inhibitor_Dose_DryPowder_Activity)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Screening Center Network, Assay Provider
BioAssay Version:
Deposit Date: 2012-12-04
Hold-until Date: 2013-09-07
Modify Date: 2013-09-10

Data Table ( Complete ):           Active    All
BioActive Compounds: 19
Depositor Specified Assays
AIDNameTypeComment
488929Elucidation of physiology of non-replicating, drug-tolerant Mycobacterium tuberculosis - Summarysummary
Description:
Keywords:
Mycobacterium tuberculosis, drug resistance, non-replicating, carbon starvation

Assay Overview:
This assay determines the inhibitory effect of small molecules against M. tuberculosis under replicating conditions

Expected Outcome:
Compounds are designated as active if 90% inhibition of survival was observed relative to DMSO control wells. All inactive compounds received an activity score of 1. The active compounds were ranked on a scale of 10-100, with 100 representing the most active compound tested in this assay.
Protocol
Medias:
1. 7H9 Complete (4.7 gm Middlebrook 7H9 powder, 900 mL water, 100 mL OADC (BD/Difco), 4 mL 50% glycerol, 2.5 mL 20% Tween-80)

Assay steps:
On the day of the assay, outside of the BSL3 a master plate containing a 2-fold dilution series of a small molecule in DMSO was created in 96 well format (top concentration 50 mM, bottom concentration DMSO only). 4uL of the master plate dilution series was transferred to the first row of an opaque 96 well assay plate. 196 uL of media was added to this row. After pipetting up and down to mix, 50 uL was transferred to the next three rows, so that the top four rows of the plate contain 50 uL of diluted small molecule in media. This process was repeated with a new small molecule in the bottom four rows of the assay plate. The assay plate was then brought into the BSL3.
Inside the BSL3, a logarithmic culture of M. tuberculosis (OD600 between 0.4-0.8) was diluted to an OD600 = 0.05 using 7H9 Complete media. 50uL of the diluted TB culture was added to each well of the 96 well plate. The plate was incubated at 37 C in a sealed Tupperware container for 7 days containing a wet kimtech shop towel. After 7 days 80uL of bacteria were removed from the 96 well plate for each concentration of drug being enumerated. The 80 uL of cells were serially diluted and plated on 7H10 agar plates. This was done in triplicate for each compound concentration being tested as well as for DMSO control wells. After 21 days the number of colonies were counted and the number of surviving bacteria calculated relative to the DMSO controls.
Comment
Result Definitions
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1Active_Concentration_QualifierString
2LogCFU_IC90*FloatμM

* Activity Concentration.
Additional Information
Grant Number: 1 R03 MH087444-01

Data Table (Concise)
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