Development of a panel of recombinase polymerase amplification assays for detection of common bacterial urinary tract infection pathogens

B. Raja, H. J. Goux, A. Marapadaga, S. Rajagopalan, K. Kourentzi, Richard C. Willson

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

Aims: To develop and evaluate the performance of a panel of isothermal real-time recombinase polymerase amplification (RPA) assays for detection of common bacterial urinary tract infection (UTI) pathogens. Methods and Results: The panel included RPAs for Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa and Enterococcus faecalis. All five RPAs required reaction times of under 12 min to reach their lower limit of detection of 100 genomes per reaction or less, and did not cross-react with high concentrations of nontarget bacterial genomic DNA. In a 50-sample retrospective clinical study, the five-RPA assay panel was found to have a specificity of 100% (95% CI, 78–100%) and a sensitivity of 89% (95% CI, 75–96%) for UTI detection. Conclusions: The analytical and clinical validity of RPA for the rapid and sensitive detection of common UTI pathogens was established. Significance and Impact of the Study: Rapid identification of the causative pathogens of UTIs can be valuable in preventing serious complications by helping avoid the empirical treatment necessitated by traditional urine culture's 48–72-h turnaround time. The routine and widespread use of RPA to supplement or replace culture-based methods could profoundly impact UTI management and the emergence of multidrug-resistant pathogens.

Original languageEnglish (US)
Pages (from-to)544-555
Number of pages12
JournalJournal of Applied Microbiology
Volume123
Issue number2
DOIs
StatePublished - Aug 2017

Keywords

  • detection
  • diagnosis
  • identification
  • rapid methods
  • urinary tract infection

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Fingerprint Dive into the research topics of 'Development of a panel of recombinase polymerase amplification assays for detection of common bacterial urinary tract infection pathogens'. Together they form a unique fingerprint.

Cite this