{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T09:56:21Z","timestamp":1753869381750,"version":"3.41.2"},"reference-count":17,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2017,5,22]],"date-time":"2017-05-22T00:00:00Z","timestamp":1495411200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81571970"],"award-info":[{"award-number":["81571970"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Clinical Laboratory Analysis"],"published-print":{"date-parts":[[2018,2]]},"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Chromoblastomycosis (CBM) is a chronic fungal disease. In China, the principle etiologic agent was a group of dematiaceous fungi, including <jats:italic>Fonsecaea monophora<\/jats:italic>,<jats:italic> Fonsecaea nubica<\/jats:italic>, and <jats:italic>Cladophialophora carrionii<\/jats:italic>. Although the <jats:italic>Fonsecaea<\/jats:italic> species have similar morphology, their pathogenicity is quite different. This study aims to establish a new solution for early identification of <jats:italic>Fonsecaea<\/jats:italic> species because of their distinctive potential infection risk.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Five reference strains and 35 clinical isolates from patients with <jats:styled-content style=\"fixed-case\">CBM<\/jats:styled-content>, preserved in our laboratory, were used in this study. The universal primer <jats:styled-content style=\"fixed-case\">ITS<\/jats:styled-content>1 and <jats:styled-content style=\"fixed-case\">ITS<\/jats:styled-content>2 were chosen to amplify the highly conserved regions of <jats:styled-content style=\"fixed-case\">rDNA<\/jats:styled-content>. High\u2010resolution melting (<jats:styled-content style=\"fixed-case\">HRM<\/jats:styled-content>) analysis was performed using the <jats:styled-content style=\"fixed-case\">LIGHTCYCLER<\/jats:styled-content><jats:sup>\u00ae<\/jats:sup> 96 System. All the amplicons were verified by direct sequencing and the sequence were aligned with those in GenBank by <jats:styled-content style=\"fixed-case\">BLAST<\/jats:styled-content> analysis.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>We successfully differentiated the five strains according to their different Tm values and curve shapes. The 35 clinical isolates from patients were identified as 24 strains for <jats:italic>F.\u00a0monophora<\/jats:italic> and 11 strains for <jats:italic>F.\u00a0nubica<\/jats:italic>, which is consistent with the <jats:styled-content style=\"fixed-case\">DNA<\/jats:styled-content> sequencing results.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>It is the first time to use <jats:styled-content style=\"fixed-case\">HRM<\/jats:styled-content> analysis for identification of <jats:italic>Fonsecaea<\/jats:italic> species. Since the CBM etiologic agent in South China is mainly <jats:italic>F.\u00a0monophora<\/jats:italic> and <jats:italic>F.\u00a0nubica<\/jats:italic>, this strategy is sufficient to be applied in the clinical examination with high accuracy, speed, and throughput.<\/jats:p><\/jats:sec>","DOI":"10.1002\/jcla.22257","type":"journal-article","created":{"date-parts":[[2017,5,23]],"date-time":"2017-05-23T23:51:56Z","timestamp":1495583516000},"source":"Crossref","is-referenced-by-count":4,"title":["High\u2010resolution melting analysis assay for identification of <i>Fonsecaea<\/i> species"],"prefix":"10.1002","volume":"32","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4307-4125","authenticated-orcid":false,"given":"Minglan","family":"Shi","sequence":"first","affiliation":[{"name":"Department of Dermatology and Venerology Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation Sun Yat\u2010sen Memorial Hospital Sun Yat\u2010sen University  Guangzhou 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Sun Yat\u2010sen University  Guangzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Liyan","family":"Xi","sequence":"additional","affiliation":[{"name":"Department of Dermatology and Venerology Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation Sun Yat\u2010sen Memorial Hospital Sun Yat\u2010sen University  Guangzhou China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Junmin","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Dermatology and Venerology Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation Sun Yat\u2010sen Memorial Hospital Sun Yat\u2010sen University  Guangzhou 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