Jiangjiang (Chris) Zhu (Presenter)
Bio: I am an assistant professor at Miami University in Ohio. The research interest of our group is developing targeted metabolic profiling approach for biological related questions, range from bacterial infection to cancer development.
Authorship: Jiangjiang (Chris) Zhu
Department of Chemistry and Biochemistry, Miami University, Oxford, Ohio 45056
Here we demonstrated a study focused on the development and application of targeted metabolic profiling for detecting and monitoring the bacterial metabolic profile changes in response to antibiotics treatment. Hydrophilic interaction liquid chromatography (HILIC) - tandem mass spectrometry method was used for metabolic profiling. More than 100 targeted metabolites, include amino acids, fatty acids, purine metabolites, pyrimidine metabolites as well as carboxylic acids have been targeted for metabolic changes due to antibiotic treatment. Our results indicated that the metabolic profile after treatment of antibiotics could aid the differentiation of MSSA strains and MRSA strains, which can be potentially applied in clinical application for antibiotic resistance test.
Antibiotics resistance has been a world-wide health threat to human health over the past decades, and the rapid detection method of bacteria antibiotic resistance is well desired in this situation. For example, Centers for Disease Control and Prevention (CDC) listed methicillin-resistant Staphylococcus aureus (MRSA) as a serious antibiotic-resistant microbial threat in the United States in 2013, which is the second-highest threat level used by the CDC. MRSA is also a growing threat in chronic lung infections, such as those associated with cystic fibrosis (CF) and chronic obstructive pulmonary disease (COPD). Genetic and molecular biology methods have been developed for many years in the effort of obtaining rapid and reliable information for bacteria antibiotics resistance test, however, the speed and outcome are not always satisfactory. Meanwhile, novel bioanalytical methods, such as mass spectrometry based metabolomics approach, has started to show promising application for providing rapid, sensitive and reproducible detection for bacteria antibiotic susceptibility.
Materials and methods:
Several strains of methicillin susceptible S. aureus (MSSA) and methicillin resistant S. aureus (MSSA) were tested in this study. All the strains were grown in both nutrition rich medium and minimal medium, and their metabolites were extracted by cold methanol extraction. Extracted metabolites were tested on the newly developed targeted metabolic profiling platform. Thermo Scientific Ultimate 3000 LC system coupled with TSQ Quantiva mass spectrometer is used in this study and the Hydrophilic interaction liquid chromatography (HILIC) - tandem mass spectrometry method was used for metabolic profiling. More than 100 metabolites have been tested using metabolite standards and then their information of retention time and selected reaction monitoring (SRM) were recorded for the targeted method setup. Multivariate statistical analysis was conducted using data analysis software JMP (SAS, INC) for the identification of significantly changed bacterial metabolites during antibiotics treatment.
Results and Discussion:
This study focused on the development and application of targeted metabolic profiling for detecting and monitoring the metabolites of bacteria during their growth, in different growth medium, and their metabolites change in response to antibiotics treatment. More than 100 targeted metabolites, include amino acids, fatty acids, purine metabolites, pyrimidine metabolites as well as carboxylic acids have been targeted for metabolic changes due to antibiotic treatment. These metabolites have been involved into studies in many important metabolic pathways from previous knowledge. Our results indicated that the metabolic profiles from S. aureus changed before and after antibiotic treatment, the measured concentration of metabolites also varied in response to different concentration of antibiotics, most importantly, the metabolic profile after treatment of antibiotics could help us distinguish MSSA strains and MRSA strains, which can be potentially useful in clinical application for antibiotic resistance test.
References & Acknowledgements:
The financial support from Miami University is gratefully acknowledged.
IP Royalty: no
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