Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Development of PCR-Multiplex Assays for Identification of the Herpotrichiellaceae Family and Agents Causing Chromoblastomycosis

Gabriel S.M. Sousa
* ORCID logo ,
Rodrigo S. De Oliveira
,
Alex B. De Souza
,
Ruan C. Monteiro
,
Elaine P. T. E. Santo
,
Luciano C. Franco Filho
,
Denison L.O. Moraes
,
Sarah R. De Sá
,
Silvia H. M. Da Silva
Version 1 : Received: 18 March 2024 / Approved: 19 March 2024 / Online: 19 March 2024 (12:35:06 CET)

How to cite: Sousa, G.S.; De Oliveira, R.S.; De Souza, A.B.; Monteiro, R.C.; Santo, E.P.T.E.; Franco Filho, L.C.; Moraes, D.L.; De Sá, S.R.; Da Silva, S.H.M. Development of PCR-Multiplex Assays for Identification of the Herpotrichiellaceae Family and Agents Causing Chromoblastomycosis. Preprints 2024, 2024031144. https://doi.org/10.20944/preprints202403.1144.v1 Sousa, G.S.; De Oliveira, R.S.; De Souza, A.B.; Monteiro, R.C.; Santo, E.P.T.E.; Franco Filho, L.C.; Moraes, D.L.; De Sá, S.R.; Da Silva, S.H.M. Development of PCR-Multiplex Assays for Identification of the Herpotrichiellaceae Family and Agents Causing Chromoblastomycosis. Preprints 2024, 2024031144. https://doi.org/10.20944/preprints202403.1144.v1

Abstract

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 aforementioned 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.

Keywords

chromoblastomycosis; phaeohyphomycosis; PCR-Multiplex; Herpotrichiellaceae; sequencing; phenotypic; molecular biology.

Subject

Biology and Life Sciences, Other

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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