{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,21]],"date-time":"2026-02-21T07:00:56Z","timestamp":1771657256082,"version":"3.50.1"},"reference-count":34,"publisher":"American Society for Microbiology","issue":"6","license":[{"start":{"date-parts":[[2017,5,24]],"date-time":"2017-05-24T00:00:00Z","timestamp":1495584000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2017,6,1]],"date-time":"2017-06-01T00:00:00Z","timestamp":1496275200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"funder":[{"name":"STRECK"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["Antimicrob Agents Chemother"],"published-print":{"date-parts":[[2017,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            <jats:named-content content-type=\"genus-species\">Escherichia coli<\/jats:named-content>\n            isolates belonging to the sequence type 131 (ST131) clonal complex have been associated with the global distribution of fluoroquinolone and \u03b2-lactam resistance. Whole-genome sequencing and multilocus sequence typing identify sequence type but are expensive when evaluating large numbers of samples. This study was designed to develop a cost-effective screening tool using high-resolution melting (HRM) analysis to differentiate ST131 from non-ST131\n            <jats:named-content content-type=\"genus-species\">E. coli<\/jats:named-content>\n            in large sample populations in the absence of sequence analysis. The method was optimized using DNA from 12\n            <jats:named-content content-type=\"genus-species\">E. coli<\/jats:named-content>\n            isolates. Singleplex PCR was performed using 10 ng of DNA, Type-it HRM buffer, and multilocus sequence typing primers and was followed by multiplex PCR. The amplicon sizes ranged from 630 to 737 bp. Melt temperature peaks were determined by performing HRM analysis at 0.1\u00b0C resolution from 50 to 95\u00b0C on a Rotor-Gene Q 5-plex HRM system. Derivative melt curves were compared between sequence types and analyzed by principal component analysis. A blinded study of 191\n            <jats:named-content content-type=\"genus-species\">E. coli<\/jats:named-content>\n            isolates of ST131 and unknown sequence types validated this methodology. This methodology returned 99.2% specificity (124 true negatives and 1 false positive) and 100% sensitivity (66 true positives and 0 false negatives). This HRM methodology distinguishes ST131 from non-ST131\n            <jats:named-content content-type=\"genus-species\">E. coli<\/jats:named-content>\n            without sequence analysis. The analysis can be accomplished in about 3 h in any laboratory with an HRM-capable instrument and principal component analysis software. Therefore, this assay is a fast and cost-effective alternative to sequencing-based ST131 identification.\n          <\/jats:p>","DOI":"10.1128\/aac.00265-17","type":"journal-article","created":{"date-parts":[[2017,4,18]],"date-time":"2017-04-18T00:55:21Z","timestamp":1492476921000},"update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":20,"title":["High-Resolution Melting Analysis for Rapid Detection of Sequence Type 131 Escherichia coli"],"prefix":"10.1128","volume":"61","author":[{"given":"Lucas B.","family":"Harrison","sequence":"first","affiliation":[{"name":"Department of Medical Microbiology and Immunology, Creighton University, Omaha, Nebraska, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nancy D.","family":"Hanson","sequence":"additional","affiliation":[{"name":"Department of Medical Microbiology and Immunology, Creighton University, Omaha, Nebraska, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_3_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.00125-13"},{"key":"e_1_3_3_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.02428-14"},{"key":"e_1_3_3_4_2","doi-asserted-by":"publisher","DOI":"10.1093\/cid\/cit503"},{"key":"e_1_3_3_5_2","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkw122"},{"key":"e_1_3_3_6_2","doi-asserted-by":"publisher","DOI":"10.1086\/669865"},{"key":"e_1_3_3_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/bs.aambs.2014.09.002"},{"key":"e_1_3_3_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.03617-13"},{"key":"e_1_3_3_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00912-13"},{"key":"e_1_3_3_10_2","first-page":"679","article-title":"Comparison of pulsed-field gel electrophoresis & repetitive sequence-based PCR methods for molecular epidemiological studies of Escherichia coli clinical isolates","volume":"140","author":"Bae IK","year":"2014","unstructured":"Bae IK KimJ SunJY JeongSH KimYR WangKK LeeK. 2014. 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