Bookmark and Share
BioAssay: AID 686977

Vibrio cholerae assay for pro-quorum sensing small molecules

Cholera is an acute water-borne diarrheal disease caused by Vibrio cholerae of serogroups O1 and O139. It has the potential to cause outbreaks wherever sanitation is poor and clean drinking water is unavailable. Recent outbreaks have occurred in sub-Saharan Africa due to political unrest and in Haiti following the2010 earthquake. Both resulted in cholera outbreaks due to a breakdown in more ..
_
   
 Tested Compounds
 Tested Compounds
All(9418)
 
 
Active(2)
 
 
Inactive(8988)
 
 
Inconclusive(428)
 
 
 Tested Substances
 Tested Substances
All(9418)
 
 
Active(2)
 
 
Inactive(8988)
 
 
Inconclusive(428)
 
 
 Related BioAssays
 Related BioAssays
AID: 686977
Data Source: Southern Research Specialized Biocontainment Screening Center (Vcholera_QS_Pilot)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
BioAssay Version:
Deposit Date: 2013-05-14
Modify Date: 2013-05-26

Data Table ( Complete ):           View Active Data    View All Data
Target
BioActive Compounds: 2
Description:
Cholera is an acute water-borne diarrheal disease caused by Vibrio cholerae of serogroups O1 and O139. It has the potential to cause outbreaks wherever sanitation is poor and clean drinking water is unavailable. Recent outbreaks have occurred in sub-Saharan Africa due to political unrest and in Haiti following the2010 earthquake. Both resulted in cholera outbreaks due to a breakdown in sanitation and water purification infrastructure. Cholera outbreaks continue to be a major public health problem in endemic areas of South Asia and Africa where cholera is estimated to cause 5 million cases and more than 100,000 deaths per year. Antibiotics are frequently used to treat cholera to mitigate the duration and severity of its life-threatening clinical symptoms in the most severe cases. This has produced antibiotic resistant strains V. cholerae O1 and O139 which can complicate treatment and limit treatment options. The development of drugs that target virulence factors, rather than viability of the bacteria, could provide novel prophylactic measures and/or adjunctive therapies to limit outbreaks and circumvent antibiotic resistance. Theoretically compounds that modulate virulence factors should not produce the kind of selection pressure that antimicrobials do and therefore should not drive the production of drug resistant strains like antibiotics do. To this end, a high throughput screen was developed to screen for compounds that enhance the quorum sensing response in V. cholera and reduce expression of virulence factors that cause disease.
Protocol
Vibrio cholerae assay for pro-quorum sensing small molecules
One hundred and twenty five nl of test compound was dispensed to a dry assay plate (Corning 3571) with an Echo555. Then 5ul of LB was added to the plates with a Matrix/Thermo Wellmate. Bacterial inoculation was also done with a Wellmate. Twenty ul of a stationary phase culture of a quorum sensing reporter strain was dispensed to the assay plates. Plates were incubated at 30C for 6 hours then read using Luminescence mode with a 0.1 second integration time on a Perkin Elmer Envision.
Comment
Possible artifacts in the V. cholerae Quorum Sensing assay include, but are not limited to, compounds that inhibit light production or kill the bacteria (false negative) and compounds that precipitate.
Compounds with luminescence values greater than 1038 (mean+3 standard deviations) were considered to show sufficient activity in the primary screen to be considered for followup. However, for this pilot study, only 2 compounds were actually tested in the confirmatory assay. Compounds showing as active in the primary screen that have no conclusive confirmatory data are categorized as "inconclusive". Primary screen data are scored based on relative activity on a scale of 0-40. Confirmatory data are scored based on reative IC50 values in the range of 41-80.
Categorized Comment - additional comments and annotations
From ChEMBL:
Assay Type: Functional
Result Definitions
Show more
TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1IC50*FloatμM
2LUM @ 100 uM Rep 1 (100μM**)Float%
3LUM @ 50 uM Rep 1 (50μM**)Float%
4LUM @ 25 uM Rep 1 (25μM**)Float%
5LUM @ 12 uM Rep 1 (12μM**)Float%
6LUM @ 6 uM Rep 1 (6μM**)Float%
7LUM @ 3 uM Rep 1 (3μM**)Float%
8LUM @ 100 uM Rep 2 (100μM**)Float%
9LUM @ 50 uM Rep 2 (50μM**)Float%
10LUM @ 25 uM Rep 2 (25μM**)Float%
11LUM @ 12 uM Rep 2 (12μM**)Float%
12LUM @ 6 uM Rep 2 (6μM**)Float%
13LUM @ 3 uM Rep 2 (3μM**)Float%
14Primary Screen LUM ValueFloat
15Primary Screen ConcentrationFloat
16Primary Screen Concentration UnitString

* Activity Concentration. ** Test Concentration.

Data Table (Concise)
Data Table ( Complete ):     View Active Data    View All Data
PageFrom: