Journal
JOURNAL OF ANTIMICROBIAL CHEMOTHERAPY
Volume 74, Issue 2, Pages 349-356Publisher
OXFORD UNIV PRESS
DOI: 10.1093/jac/dky419
Keywords
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Funding
- University of East Anglia
- BBSRC [BBS/E/F/000PR10348] Funding Source: UKRI
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Background: Increasing resistance drives empirical use of less potent and previously reserved antibiotics, including for urinary tract infections (UTIs). Molecular profiling, without culture, might better guide early therapy. Objectives: To explore the potential of AusDiagnostics multiplex tandem (MT) PCR UTI assays. Methods: Two MT-PER assays were developed successively, seeking 8 or 16 resistance genes. Amplification was tracked in real time, with melting temperatures used to confirm product identity. Assays were variously performed on: (i) extracted DNA; (ii) cultured bacteria; (iii) urine spiked with reference strains; and (iv) bacteria harvested from clinical urines. Results were compared with those from sequencing, real-time SybrGreen PER or phenotypic susceptibility. Results: Performance was similar irrespective of whether DNA, cultures or urines were used, with >90% sensitivity and specificity with respect to common fi-lactamases, dfr genes and a minoglycoside resistance determinants except aadA1/42/A3, for which carriage correlated poorly with streptomycin resistance. Fluoroquinolonesusceptible and-resistant Escherichia coil (but not other species) were distinguished by the melting temperatures of their gyrA PCR products. The time from urine to results was <3 h. Conclusions: The MT-PCR assays rapidly identified resistance genes from Gram-negative bacteria in urines as well as from cultivated bacteria. Used directly on urines, this assay has the potential to guide early therapy.
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