title: An Automated Miniaturized Method to Perform and Analyze Antimicrobial Drug Synergy Assays.
abstract: In the light of emerging antibiotic resistance mechanisms found in bacteria throughout the world, discovery of drugs that potentiate the effect of currently available antibiotics remains an important aspect of pharmaceutical research in the 21st century. Well-established clinical tests exist to determine synergy in vitro, but these are only optimal for low-throughput experimentation while leaving analysis of results and interpretation of high-throughput microscale assays poorly standardized. Here, we describe a miniaturized broth microdilution checkerboard assay and data analysis method in 384-well plate format that conforms to the Clinical Laboratory and Standards Institute (CLSI) methods. This method has been automated and developed to rapidly determine the synergism of current antibiotics with various beta-lactamase inhibitors emerging from our antimicrobial research efforts. This technique increases test throughput and integrity of results, and saves test compound and labor. We facilitated the interpretation of results with an automated analysis tool allowing us to rapidly qualify inter- and intraplate robustness, determine efficacy of multiple antibiotics at the same time, and standardize the results of synergy interpretation. This procedure should enhance high-throughput antimicrobial drug discovery and supersedes former techniques.
authors.affiliation: Lead Identification, Department of Molecular Therapeutics, The Scripps Research Institute , Scripps Florida, Jupiter, Florida.
authors.forename: Peter
authors.initials: P
authors.lastname: Chase
authors.affiliation: Lead Identification, Department of Molecular Therapeutics, The Scripps Research Institute , Scripps Florida, Jupiter, Florida.
authors.forename: Imarhia
authors.initials: I
authors.lastname: Enogieru
authors.affiliation: Lead Identification, Department of Molecular Therapeutics, The Scripps Research Institute , Scripps Florida, Jupiter, Florida.
authors.forename: Franck
authors.initials: F
authors.lastname: Madoux
authors.affiliation: Lead Identification, Department of Molecular Therapeutics, The Scripps Research Institute , Scripps Florida, Jupiter, Florida.
authors.forename: Eric
authors.initials: E
authors.lastname: Bishop
authors.affiliation: Lead Identification, Department of Molecular Therapeutics, The Scripps Research Institute , Scripps Florida, Jupiter, Florida.
authors.forename: Jacob
authors.initials: J
authors.lastname: Beer
authors.affiliation: Lead Identification, Department of Molecular Therapeutics, The Scripps Research Institute , Scripps Florida, Jupiter, Florida.
authors.forename: Louis
authors.initials: L
authors.lastname: Scampavia
authors.affiliation: Lead Identification, Department of Molecular Therapeutics, The Scripps Research Institute , Scripps Florida, Jupiter, Florida.
authors.forename: Timothy
authors.initials: T
authors.lastname: Spicer
mesh_terms: D000890:Anti-Infective Agents / Q000032:analysis*; D004357:Drug Synergism; D008826:Microbial Sensitivity Tests / Q000379:methods*; D008904:Miniaturization / Q000379:methods*
chemical_list: D000890:Anti-Infective Agents