Energy management of Uropathogenic Escherichia coli during Trimethoprim challenge for the Rapid Antimicrobial susceptibility testing of urinary tract infections

Carter, Cailean Andrew (2023) Energy management of Uropathogenic Escherichia coli during Trimethoprim challenge for the Rapid Antimicrobial susceptibility testing of urinary tract infections. Doctoral thesis, University of East Anglia.

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Abstract

Introduction: There is a long history behind ATP bioluminescence as a cost-effective and rapid means of detecting bacteria and antibiotic susceptibility for clinical use. Treatments for common illnesses like urinary tract infections (UTIs) rely on empirical or delayed antibiotic treatments and would benefit greatly from technology like ATP bioluminescence. However, the challenge has been achieving rapid susceptibility results for all clinically used antibiotics like trimethoprim.

Aim: To devise a way of detecting trimethoprim susceptibility using ATP bioluminescence within 30 minutes and determining the causal mechanism of ATP depletion during trimethoprim challenge.

Methods: Two hundred and seventeen clinical Uropathogenic E. coli isolates were collected with resistance data and whole genome sequenced for comparative genomics, identifying resistance mechanisms, and key groups causing UTIs locally. Isolates were screened using ATP bioluminescence with different metabolites to identify conditions suitable for rapid trimethoprim susceptibility detection. Flux balance analysis (FBA) was performed on a genome-scale metabolic model of E. coli UTI89 incorporating in vivo ATP data to identify the role of various media components and ATP during trimethoprim challenge.

Results: Purines were identified as a prohibitive factor in rapid trimethoprim susceptibility testing. By excluding purines, trimethoprim susceptibility results could be achieved in 30 minutes of exposure. FBA suggested free ATP was salvaged to restore trimethoprim-induced purine and nucleoside depletion needed for DNA and RNA synthesis.

Conclusion: This work provides novel insights into how bacterial metabolism can be manipulated to achieve rapid trimethoprim susceptibility testing with potential applications in point-of-care testing for UTIs.

Item Type:	Thesis (Doctoral)
Faculty \ School:	Faculty of Medicine and Health Sciences > Norwich Medical School
Depositing User:	Nicola Veasy
Date Deposited:	24 Jan 2024 15:22
Last Modified:	24 Jan 2024 15:22
URI:	https://ueaeprints.uea.ac.uk/id/eprint/94243
DOI:

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