Whole Animal Automated Platform for Drug Discovery against Multi-Drug Resistant Staphylococcus aureus
View/ Open
Author
Rajamuthiah, Rajmohan
Fuchs, Beth Burgwyn
Kim, Younghoon
Larkins-Ford, Jonah
Published Version
https://doi.org/10.1371/journal.pone.0089189Metadata
Show full item recordCitation
Rajamuthiah, Rajmohan, Beth Burgwyn Fuchs, Elamparithi Jayamani, Younghoon Kim, Jonah Larkins-Ford, Annie Conery, Frederick M. Ausubel, and Eleftherios Mylonakis. 2014. “Whole Animal Automated Platform for Drug Discovery against Multi-Drug Resistant Staphylococcus aureus.” PLoS ONE 9 (2): e89189. doi:10.1371/journal.pone.0089189. http://dx.doi.org/10.1371/journal.pone.0089189.Abstract
Staphylococcus aureus, the leading cause of hospital-acquired infections in the United States, is also pathogenic to the model nematode Caenorhabditis elegans. The C. elegans-S. aureus infection model was previously carried out on solid agar plates where the bacteriovorous C. elegans feeds on a lawn of S. aureus. However, agar-based assays are not amenable to large scale screens for antibacterial compounds. We have developed a high throughput liquid screening assay that uses robotic instrumentation to dispense a precise amount of methicillin resistant S. aureus (MRSA) and worms in 384-well assay plates, followed by automated microscopy and image analysis. In validation of the liquid assay, an MRSA cell wall defective mutant, MW2ΔtarO, which is attenuated for killing in the agar-based assay, was found to be less virulent in the liquid assay. This robust assay with a Z’-factor consistently greater than 0.5 was utilized to screen the Biomol 4 compound library consisting of 640 small molecules with well characterized bioactivities. As proof of principle, 27 of the 30 clinically used antibiotics present in the library conferred increased C. elegans survival and were identified as hits in the screen. Surprisingly, the antihelminthic drug closantel was also identified as a hit in the screen. In further studies, we confirmed the anti-staphylococcal activity of closantel against vancomycin-resistant S. aureus isolates and other Gram-positive bacteria. The liquid C. elegans – S. aureus assay described here allows screening for anti-staphylococcal compounds that are not toxic to the host.Other Sources
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3929655/pdf/Terms of Use
This article is made available under the terms and conditions applicable to Other Posted Material, as set forth at http://nrs.harvard.edu/urn-3:HUL.InstRepos:dash.current.terms-of-use#LAACitable link to this page
http://nrs.harvard.edu/urn-3:HUL.InstRepos:11879738
Collections
- FAS Scholarly Articles [18292]
- HMS Scholarly Articles [17922]
Contact administrator regarding this item (to report mistakes or request changes)