{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T06:20:16Z","timestamp":1791958816628,"version":"4.3.4"},"reference-count":49,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2024,8,4]],"date-time":"2024-08-04T00:00:00Z","timestamp":1722729600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Institute Evandro Chagas (IEC), Department of Health Surveillance, Ministry of Health: Laboratory of Superficial and Systemic Mycoses","award":["CAPES\u201488887.840006\/2023-00"],"award-info":[{"award-number":["CAPES\u201488887.840006\/2023-00"]}]},{"name":"Coordena\u00e7\u00e3o de Aperfei\u00e7oamento de Pessoal de N\u00edvel Superior\u2014Brasil","award":["CAPES\u201488887.840006\/2023-00"],"award-info":[{"award-number":["CAPES\u201488887.840006\/2023-00"]}]},{"name":"PROPESP\/UFPA (PAPQ)","award":["CAPES\u201488887.840006\/2023-00"],"award-info":[{"award-number":["CAPES\u201488887.840006\/2023-00"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["JoF"],"abstract":"<jats:p>The Herpotrichiellaceae family is an important group of dematiaceous filamentous fungi, associated with a variety of pathogenic fungal species causing chromoblastomycosis (CBM) and phaeohyphomycosis (PHM), both with polymorphic clinical manifestations and worldwide incidence. Currently, the identification of this family and determination of the causative agent is challenging due to the subjectivity of morphological identification methods, necessitating the use of molecular techniques to complement diagnosis. In this context, genetic sequencing of the Internal Transcribed Spacer (ITS) has become the norm due to a lack of alternative molecular tools for identifying these agents. Therefore, this study aimed to develop PCR-Multiplex methodologies to address this gap. Sequences from the ITS and Large Subunit (LSU) of ribosomal DNA were used, and after manual curation and in vitro analyses, primers were synthesized for the identification of the targets. The primers were optimized and validated in vitro, resulting in two PCR-Multiplex methodologies: one for identifying the Herpotrichiellaceae family and the bantiana clade, and another for determining the species Fonsecaea pedrosoi and Fonsecaea monophora. Ultimately, the assays developed in this study aim to complement other identification approaches for these agents, reducing the need for sequencing, improving the management of these infections, and enhancing the accuracy of epidemiological information.<\/jats:p>","DOI":"10.3390\/jof10080548","type":"journal-article","created":{"date-parts":[[2024,8,5]],"date-time":"2024-08-05T13:57:28Z","timestamp":1722866248000},"page":"548","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Development of PCR-Multiplex Assays for Identification of the Herpotrichiellaceae Family and Agents Causing Chromoblastomycosis"],"prefix":"10.3390","volume":"10","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2605-3755","authenticated-orcid":false,"given":"Gabriel S. M.","family":"Sousa","sequence":"first","affiliation":[{"name":"Programa de P\u00f3s-Gradua\u00e7\u00e3o em Biologia de Agentes Infecciosos e Parasit\u00e1rios, Instituto de Ci\u00eancias Biol\u00f3gicas, Universidade Federal do Par\u00e1, Bel\u00e9m 66075-750, Brazil"},{"name":"Laborat\u00f3rio de Micoses Superficiais e Sist\u00eamicas, Se\u00e7\u00e3o de Bacteriologia e Micologia, Instituto Evandro Chagas, Ananindeua 67030-000, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rodrigo S.","family":"De Oliveira","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Micoses Superficiais e Sist\u00eamicas, Se\u00e7\u00e3o de Bacteriologia e Micologia, Instituto Evandro Chagas, Ananindeua 67030-000, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1244-4026","authenticated-orcid":false,"given":"Alex B.","family":"Souza","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Micoses Superficiais e Sist\u00eamicas, Se\u00e7\u00e3o de Bacteriologia e Micologia, Instituto Evandro Chagas, Ananindeua 67030-000, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8246-5543","authenticated-orcid":false,"given":"Ruan C.","family":"Monteiro","sequence":"additional","affiliation":[{"name":"Laboratory of Emerging Fungal Pathogens, Universidade Federal de S\u00e3o Paulo, S\u00e3o Paulo 04023-062, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elaine P. T. E.","family":"Santo","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Micoses Superficiais e Sist\u00eamicas, Se\u00e7\u00e3o de Bacteriologia e Micologia, Instituto Evandro Chagas, Ananindeua 67030-000, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5648-4118","authenticated-orcid":false,"given":"Luciano C.","family":"Franco Filho","sequence":"additional","affiliation":[{"name":"Laborat\u00f3rio de Micoses Superficiais e Sist\u00eamicas, Se\u00e7\u00e3o de Bacteriologia e Micologia, Instituto Evandro Chagas, Ananindeua 67030-000, Brazil"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Denison L. 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