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The MYCO WELL D-ONE is a culture-based assay that aims to detect these organisms whilst concurrently screening them for antibiotic resistance. Urine and\/or swabs from 856 informed and consented participants attending Welsh sexual health clinics were subjected to MYCO WELL D-ONE analysis, alongside qPCR and culture titration methodologies to determine sensitivity, specificity, PPV, NPV and accuracy. Resistance was confirmed by CLSI-compliant susceptibility testing and genetic mechanisms determined. The MYCO WELL D-ONE displayed a sensitivity and specificity of 91.98% and 96.44% for the detection of <jats:italic>Ureaplasma<\/jats:italic> spp., with sensitivity and specificity values of 78.23% and 98.84% for <jats:italic>Mycoplasma hominis<\/jats:italic>, compared with qPCR. Swabs harboured significantly greater bacterial loads than urine samples for both <jats:italic>Ureaplasma<\/jats:italic> spp. and <jats:italic>M<\/jats:italic>. <jats:italic>hominis<\/jats:italic>. Levofloxacin resistance rates, mediated by Ser83Leu mutation in ParC, for <jats:italic>Ureaplasma<\/jats:italic> spp. were 0.54%. Tetracycline resistance rates, mediated by <jats:italic>tet<\/jats:italic>(M), were 0.54% and 2% for <jats:italic>Ureaplasma<\/jats:italic> spp. and <jats:italic>M<\/jats:italic>. <jats:italic>hominis<\/jats:italic>, respectively; sequence analysis of <jats:italic>tet<\/jats:italic>(M)-positive <jats:italic>Ureaplasma<\/jats:italic> spp. and <jats:italic>M<\/jats:italic>. <jats:italic>hominis<\/jats:italic> strains isolated from a single individual confirmed separate resistance gene origins. The MYCO WELL D-ONE is a sensitive and specific assay for the detection of <jats:italic>Ureaplasma<\/jats:italic> spp. and <jats:italic>M<\/jats:italic>. <jats:italic>hominis<\/jats:italic> in genitourinary medicine samples, facilitating the accurate detection of these organisms within low-technology environments. While good for antibiotic resistance screening, accurate confirmation by MIC determination or molecular methods are required, and more optimally performed on urine samples.<\/jats:p>","DOI":"10.1007\/s10096-020-03993-7","type":"journal-article","created":{"date-parts":[[2020,7,28]],"date-time":"2020-07-28T17:26:17Z","timestamp":1595957177000},"page":"2427-2440","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":22,"title":["MYCO WELL D-ONE detection of Ureaplasma spp. and Mycoplasma hominis in sexual health patients in Wales"],"prefix":"10.1007","volume":"39","author":[{"given":"Daniel J.","family":"Morris","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lucy C.","family":"Jones","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rebecca L.","family":"Davies","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kirsty","family":"Sands","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Edward","family":"Portal","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9117-6911","authenticated-orcid":false,"given":"Owen B.","family":"Spiller","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2020,7,28]]},"reference":[{"key":"3993_CR1","unstructured":"Bhimji SS, Unakal CG (2017) Bacteria, Gram Positive. 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