title: Monitoring bacterial resistance to chloramphenicol and other antibiotics by liquid chromatography electrospray ionization tandem mass spectrometry using selected reaction monitoring.
abstract: Antibiotic resistance is a growing problem worldwide. For this reason, clinical laboratories often determine the susceptibility of the bacterial isolate to a number of different antibiotics in order to establish the most effective antibiotic for treatment. Unfortunately, current susceptibility assays are time consuming. Antibiotic resistance often involves the chemical modification of an antibiotic to an inactive form by an enzyme expressed by the bacterium. Selected reaction monitoring (SRM) has the ability to quickly monitor and identify these chemical changes in an unprecedented time scale. In this work, we used SRM as a technique to determine the susceptibility of several different antibiotics to the chemically modifying enzymes β-lactamase and chloramphenicol acetyltransferase, enzymes used by bacteria to confer resistance to major classes of commonly used antibiotics. We also used this technique to directly monitor the effects of resistant bacteria grown in a broth containing a specific antibiotic. Because SRM is highly selective and can also identify chemical changes in a multitude of antibiotics in a single assay, SRM has the ability to detect organisms that are resistant to multiple antibiotics in a single assay. For these reasons, the use of SRM greatly reduces the time it takes to determine the susceptibility or resistance of an organism to a multitude of antibiotics by eliminating the time-consuming process found in other currently used methods.
authors.affiliation: Biomolecular Resource Facility, University of Texas Medical Branch, Galveston, TX 77555, USA. amhaag@utmb.edu
authors.forename: Anthony M
authors.initials: AM
authors.lastname: Haag
authors.forename: Audrie M
authors.initials: AM
authors.lastname: Medina
authors.forename: Ariel E
authors.initials: AE
authors.lastname: Royall
authors.forename: Norbert K
authors.initials: NK
authors.lastname: Herzog
authors.forename: David W
authors.initials: DW
authors.lastname: Niesel
mesh_terms: D000900:Anti-Bacterial Agents / Q000032:analysis* / Q000737:chemistry / Q000494:pharmacology*; D002701:Chloramphenicol / Q000032:analysis* / Q000737:chemistry / Q000494:pharmacology*; D002853:Chromatography, Liquid / Q000379:methods*; D024881:Drug Resistance, Bacterial; D004926:Escherichia coli / Q000737:chemistry / Q000187:drug effects; D016014:Linear Models; D008954:Models, Biological; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry
chemical_list: D000900:Anti-Bacterial Agents; D002701:Chloramphenicol