{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,3,8]],"date-time":"2025-03-08T05:23:53Z","timestamp":1741411433341,"version":"3.38.0"},"reference-count":38,"publisher":"American Veterinary Medical Association (AVMA)","issue":"7","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["ajvr"],"published-print":{"date-parts":[[2018,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n<jats:p><jats:bold>OBJECTIVE<\/jats:bold> To evaluate a fluorescence resonance energy transfer quantitative PCR (FRET-qPCR) assay for detection of <jats:italic>gyrA<\/jats:italic> mutations conferring fluoroquinolone resistance in canine urinary <jats:italic>Escherichia coli<\/jats:italic> isolates and canine urine specimens.<\/jats:p>\n<jats:p><jats:bold>SAMPLE<\/jats:bold> 264 canine urinary <jats:italic>E coli<\/jats:italic> isolates and 283 clinical canine urine specimens.<\/jats:p>\n<jats:p><jats:bold>PROCEDURES<\/jats:bold> The <jats:italic>E coli<\/jats:italic> isolates were used to validate the FRET-qPCR assay. Urine specimens were evaluated by bacterial culture and identification, isolate enrofloxacin susceptibility testing, and FRET-qPCR assay. Sensitivity and specificity of the FRET-qPCR assay for detection of <jats:italic>gyrA<\/jats:italic> mutations in urine specimens and in <jats:italic>E coli<\/jats:italic> isolated from urine specimens were computed, with results of enrofloxacin susceptibility testing used as the reference standard.<\/jats:p>\n<jats:p><jats:bold>RESULTS<\/jats:bold> The validated FRET-qPCR assay discriminated between enrofloxacin-resistant and enrofloxacin-susceptible <jats:italic>E coli<\/jats:italic> isolates with an area under the receiver operating characteristic curve of 0.92. The assay accurately identified 25 of 40 urine specimens as containing enrofloxacin-resistant isolates (sensitivity, 62.5%) and 226 of 243 urine specimens as containing enrofloxacin-susceptible isolates (specificity, 93.0%). When the same assay was performed on <jats:italic>E coli<\/jats:italic> isolates recovered from these specimens, sensitivity (77.8%) and specificity (94.8%) increased. Moderate agreement was achieved between results of the FRET-qPCR assay and enrofloxacin susceptibility testing for <jats:italic>E coli<\/jats:italic> isolates recovered from urine specimens.<\/jats:p>\n<jats:p><jats:bold>CONCLUSIONS AND CLINICAL RELEVANCE<\/jats:bold> The FRET-qPCR assay was able to rapidly distinguish between enrofloxacin-resistant and enrofloxacin-susceptible <jats:italic>E coli<\/jats:italic> in canine clinical urine specimens through detection of <jats:italic>gyrA<\/jats:italic> mutations. Therefore, the assay may be useful in clinical settings to screen such specimens for enrofloxacin-resistant <jats:italic>E coli<\/jats:italic> to avoid inappropriate use of enrofloxacin and contributing to antimicrobial resistance.<\/jats:p>","DOI":"10.2460\/ajvr.79.7.755","type":"journal-article","created":{"date-parts":[[2018,6,26]],"date-time":"2018-06-26T12:19:37Z","timestamp":1530015577000},"page":"755-761","source":"Crossref","is-referenced-by-count":0,"title":["Evaluation of a fluorescence resonance energy transfer quantitative polymerase chain reaction assay for identification of gyrA mutations conferring enrofloxacin resistance in canine urinary Escherichia coli isolates and canine urine specimens"],"prefix":"10.2460","volume":"79","author":[{"given":"Megan G.","family":"Behringer","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dawn 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