The development and evaluation of a multiplex real-time PCR assay for the detection of ESBL genes in urinary tract infections

Date

2018-08-09

Advisors

Journal Title

Journal ISSN

ISSN

Volume Title

Publisher

Open Access Pub

Type

Article

Peer reviewed

Yes

Abstract

Background Overuse of beta-lactam antibiotics has lead to selection for extended-spectrum β-lactamase (ESBL) producing Enterobacteriaceae, a major cause of antibiotic resistant urinary tract infections (UTIs). Standard detection methods are time-consuming, with disputed accuracy.
This study describes a novel real-time PCR method to detect CTX-M, SHV, OXA and TEM. Methods 179 Enterobacteriaceae isolates from UTIs were collected from the Leicester Royal Infirmary, UK. A multiplex Plexor®-based real-time PCR assay detected ESBLs using their specific amplicon melting temperature, during each cycle, removing the need for a melt-curve analysis. Validation was achieved by end-point PCR and disk diffusion. Results The method was able to produce rapid and accurate results, achieving a sensitivity and specificity of 94.9% and 72% respectively, and the assay can differentiate between the different ESBL genes, with ease. Conclusions With further investigation, a Plexor®-based assay could form the basis of a high-throughput kit that health services could use to detect ESBLs or other antibiotic resistance genes.

Description

open access journal

Keywords

Extended-spectrum beta-lactamases, urinary tract infections, Real-Time PCR, rapid diagnostics, antibiotic resistance

Citation

Reid, R. and Samarasinghe, S. (2018) The development and evaluation of a multiplex real-time PCR assay for the detection of ESBL genes in urinary tract infections. International Journal of Clinical Microbiology, 1 (1), pp. 16-24

Rights

Research Institute

Leicester Institute for Pharmaceutical Innovation - From Molecules to Practice (LIPI)