Staphylococcus aureus methicillin resistance detected by HPLC-MS/MS targeted metabolic profiling.
Recently, novel bioanalytical methods, such as NMR and mass spectrometry based metabolomics approaches, have started to show promise in providing rapid, sensitive and reproducible detection of Staphylococcus aureus antibiotic resistance. Here we performed a proof-of-concept study focused on the application of HPLC-MS/MS based targeted metabolic profiling for detecting and monitoring the bacterial metabolic profile changes in response to sub-lethal levels of methicillin exposure. One hundred seventy-seven targeted metabolites from over 20 metabolic pathways were specifically screened and one hundred and thirty metabolites from in vitro bacterial tests were confidently detected from both methicillin susceptible and methicillin resistant Staphylococcus aureus (MSSA and MRSA, respectively). The metabolic profiles can be used to distinguish the isogenic pairs of MSSA strains from MRSA strains, without or with sub-lethal levels of methicillin exposure. In addition, better separation between MSSA and MRSA strains can be achieved in the latter case using principal component analysis (PCA). Metabolite data from isogenic pairs of MSSA and MRSA strains were further compared without and with sub-lethal levels of methicillin exposure, with metabolic pathway analyses additionally performed. Both analyses suggested that the metabolic activities of MSSA strains were more susceptible to the perturbation of the sub-lethal levels of methicillin exposure compared to the MRSA strains.
Department of Chemistry and Biochemistry, Miami University, 651 E High St., Oxford, OH 45056, USA.
Katie
K
Schelli
Department of Chemistry and Biochemistry, Miami University, 651 E High St., Oxford, OH 45056, USA.
Joshua
J
Rutowski
Department of Chemistry and Biochemistry, Miami University, 651 E High St., Oxford, OH 45056, USA.
Julia
J
Roubidoux
Department of Chemistry and Biochemistry, Miami University, 651 E High St., Oxford, OH 45056, USA. Electronic address: zhuj6@miamioh.edu.
Jiangjiang
J
Zhu
D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006801:Humans; D053858:Metabolic Networks and Pathways / Q000187:drug effects; D055442:Metabolome / Q000187:drug effects*; D055432:Metabolomics / Q000379:methods; D008712:Methicillin / Q000494:pharmacology*; D016106:Methicillin Resistance; D055624:Methicillin-Resistant Staphylococcus aureus / Q000187:drug effects* / Q000302:isolation & purification / Q000378:metabolism*; D013203:Staphylococcal Infections / Q000188:drug therapy / Q000382:microbiology*; D013211:Staphylococcus aureus / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism; D053719:Tandem Mass Spectrometry / Q000379:methods
D000900:Anti-Bacterial Agents; D008712:Methicillin
Antibiotic resistance test; Mass spectrometry based metabolomics; Metabolic pathway analysis; Methicillin resistant Staphylococcus aureus; Targeted metabolic profiling