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

dc.cclicenceCC-BYen
dc.contributor.authorReid, R.en
dc.contributor.authorSamarasinghe, S.en
dc.date.acceptance2018-08-08en
dc.date.accessioned2018-08-30T12:42:30Z
dc.date.available2018-08-30T12:42:30Z
dc.date.issued2018-08-09
dc.descriptionopen access journalen
dc.description.abstractBackground 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.en
dc.funderN/Aen
dc.identifier.citationReid, 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. 16-24en
dc.identifier.doihttps://doi.org/10.14302/issn.2690-4721.ijcm-18-2217
dc.identifier.urihttps://oap-lifescience.org/ijcm/article/808
dc.identifier.urihttp://hdl.handle.net/2086/16517
dc.language.isoenen
dc.peerreviewedYesen
dc.projectidN/Aen
dc.publisherOpen Access Puben
dc.researchgroupInfectious Disease Research Groupen
dc.researchinstituteLeicester Institute for Pharmaceutical Innovation - From Molecules to Practice (LIPI)en
dc.subjectExtended-spectrum beta-lactamasesen
dc.subjecturinary tract infectionsen
dc.subjectReal-Time PCRen
dc.subjectrapid diagnosticsen
dc.subjectantibiotic resistanceen
dc.titleThe development and evaluation of a multiplex real-time PCR assay for the detection of ESBL genes in urinary tract infectionsen
dc.typeArticleen

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