Malaria mitochondrial diagnosis: Challenges and Pitfalls.

Detalhes bibliográficos
Autor(a) principal: Costa, Gabriel Luíz
Data de Publicação: 2023
Outros Autores: Alvarenga, Denise Anete Madureira de, Assis, Gabriela Maíra Pereira de, Aguiar, Anna Caroline Campos, Louzada, Jaime, Pereira, Dhélio Batista, Pina-Costa, Anielle de, Hirano, Zelinda Maria Braga, Moreira, Sílvia Bahadian, Pissinatti, Alcides, Brasil, Patrícia, Daniel-Ribeiro, Cláudio Tadeu, Sousa, Taís Nóbrega de, Brito, Cristiana Ferreira Alves de
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/60050
Resumo: Fundação Oswaldo Cruz. Instituto René Rachou. Grupo de Pesquisas em Biologia Molecular e Imunologia de Malária. Belo Horizonte, MG, Brazil
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spelling Costa, Gabriel LuízAlvarenga, Denise Anete Madureira deAssis, Gabriela Maíra Pereira deAguiar, Anna Caroline CamposLouzada, JaimePereira, Dhélio BatistaPina-Costa, Anielle deHirano, Zelinda Maria BragaMoreira, Sílvia BahadianPissinatti, AlcidesBrasil, PatríciaDaniel-Ribeiro, Cláudio TadeuSousa, Taís Nóbrega deBrito, Cristiana Ferreira Alves de2023-08-11T18:49:39Z2023-08-11T18:49:39Z2023COSTA, Gabriel Luiz et al. Malaria mitochondrial diagnosis: Challenges and Pitfalls. Frontiers in Tropical Diseases, v. 4, p. 1204195, doi.org/10.3389/fitd.2023.1204195.2673-7515https://www.arca.fiocruz.br/handle/icict/60050engFrontiers Media S.A.Malaria mitochondrial diagnosis: Challenges and Pitfalls.info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto René Rachou. Grupo de Pesquisas em Biologia Molecular e Imunologia de Malária. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Instituto René Rachou. Grupo de Pesquisas em Biologia Molecular e Imunologia de Malária. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Instituto René Rachou. Grupo de Pesquisas em Biologia Molecular e Imunologia de Malária. Belo Horizonte, MG, BrazilUniversidade Federal de São Paulo. Departamento de Biociência. Santos, SP, BrazilUniversidade Federal de Roraima. Laboratório de Monitoramento de Artrópodes Vetores da Amazônia. Boa Vista, RO, BrazilCentro de Pesquisas em Medicina Tropical. Porto Velho, TO, BrazilFundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Doenças Febris Agudas. Rio de Janeiro, RJ, Brazil/Fundação Oswaldo Cruz. Centro de Pesquisa, Diagnóstico e Treinamento em Malária. Rio de Janeiro, RJ, Brazil/Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Malária. Rio de Janeiro, RJ, Brazil/Universidade Federal Fluminense. Departamento de Doenças Infecciosas e parasitárias. Escola de Enfermagem Aurora de Afonso Costa. Niterói, RJ, BrazilCentro de Pesquisas Biológicas de Indaial. Indaial, SP, Brazil/Universidade Regional de Blumenal. Blumenau, SC, BrazilInstituto Estadual do Ambiente. Centro de Primatologia do Rio de Janeiro. Guapimirim, RJ, BrazilInstituto Estadual do Ambiente. Centro de Primatologia do Rio de Janeiro. Guapimirim, RJ, Brazil/Centro Universitário Serra dos Órgãos. Teresópolis, RJ, BrazilFundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Doenças Febris Agudas. Rio de Janeiro, RJ, BrazilFundação Oswaldo Cruz. Centro de Pesquisa, Diagnóstico e Treinamento em Malária. Rio de Janeiro, RJ, Brazil/Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Malária. Rio de Janeiro, RJ, BrazilFundação Oswaldo Cruz. Instituto René Rachou. Grupo de Pesquisas em Biologia Molecular e Imunologia de Malária. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Instituto René Rachou. Grupo de Pesquisas em Biologia Molecular e Imunologia de Malária. Belo Horizonte, MG, BrazilBackground: High-copy genomic sequences could be used as PCR targets for the detection of Plasmodium infections, providing increased sensitivity over single- or low-copy genes. Mitochondrial genomes of malaria parasites are present in multiple copies in a single mitochondrion, and each parasite has many mitochondria. Here, we describe the development of seven species-specific qPCR assays for the diagnosis of Plasmodium vivax and Plasmodium falciparum, targeting coding and non-coding mitochondrial genomic regions. Methods: The optimization of the qPCR protocols involved a gradient of annealing temperatures and concentrations of primers and probes, as well as the inclusion of PCR additives/enhancers [e.g., dimethyl sulfoxide (DMSO), glycerol, bovine serum albumin (BSA)] to improve the specificity of qPCR amplification. Results: Non-specific amplification of other Plasmodium species and of human targets was observed in different levels for all assays. Regardless of the late Cq values for most non-specific amplifications, the application of a cutoff value did not completely exclude false-positive amplification, compromising the specificity and also the sensitivity of the assays. Conclusions: Therefore, although mitochondrial targets have higher sensitivity, they frequently lose specificity due to their high levels of sequence conservation. A screening to evaluate the cross-reaction between Plasmodium species and the non-specific amplification of human malaria-free samples must be performed for Plasmodium mitochondrial assays.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/60050/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALMalaria mitochondrial diagnosis challenges and pitfalls - fitd-04-1204195.pdfMalaria mitochondrial diagnosis challenges and pitfalls - fitd-04-1204195.pdfapplication/pdf489585https://www.arca.fiocruz.br/bitstream/icict/60050/2/Malaria%20mitochondrial%20diagnosis%20challenges%20and%20pitfalls%20-%20fitd-04-1204195.pdf856a4784cf25c3b7cc38ea0444e1f5baMD52icict/600502023-08-11 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dc.title.en_US.fl_str_mv Malaria mitochondrial diagnosis: Challenges and Pitfalls.
title Malaria mitochondrial diagnosis: Challenges and Pitfalls.
spellingShingle Malaria mitochondrial diagnosis: Challenges and Pitfalls.
Costa, Gabriel Luíz
title_short Malaria mitochondrial diagnosis: Challenges and Pitfalls.
title_full Malaria mitochondrial diagnosis: Challenges and Pitfalls.
title_fullStr Malaria mitochondrial diagnosis: Challenges and Pitfalls.
title_full_unstemmed Malaria mitochondrial diagnosis: Challenges and Pitfalls.
title_sort Malaria mitochondrial diagnosis: Challenges and Pitfalls.
author Costa, Gabriel Luíz
author_facet Costa, Gabriel Luíz
Alvarenga, Denise Anete Madureira de
Assis, Gabriela Maíra Pereira de
Aguiar, Anna Caroline Campos
Louzada, Jaime
Pereira, Dhélio Batista
Pina-Costa, Anielle de
Hirano, Zelinda Maria Braga
Moreira, Sílvia Bahadian
Pissinatti, Alcides
Brasil, Patrícia
Daniel-Ribeiro, Cláudio Tadeu
Sousa, Taís Nóbrega de
Brito, Cristiana Ferreira Alves de
author_role author
author2 Alvarenga, Denise Anete Madureira de
Assis, Gabriela Maíra Pereira de
Aguiar, Anna Caroline Campos
Louzada, Jaime
Pereira, Dhélio Batista
Pina-Costa, Anielle de
Hirano, Zelinda Maria Braga
Moreira, Sílvia Bahadian
Pissinatti, Alcides
Brasil, Patrícia
Daniel-Ribeiro, Cláudio Tadeu
Sousa, Taís Nóbrega de
Brito, Cristiana Ferreira Alves de
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Costa, Gabriel Luíz
Alvarenga, Denise Anete Madureira de
Assis, Gabriela Maíra Pereira de
Aguiar, Anna Caroline Campos
Louzada, Jaime
Pereira, Dhélio Batista
Pina-Costa, Anielle de
Hirano, Zelinda Maria Braga
Moreira, Sílvia Bahadian
Pissinatti, Alcides
Brasil, Patrícia
Daniel-Ribeiro, Cláudio Tadeu
Sousa, Taís Nóbrega de
Brito, Cristiana Ferreira Alves de
description Fundação Oswaldo Cruz. Instituto René Rachou. Grupo de Pesquisas em Biologia Molecular e Imunologia de Malária. Belo Horizonte, MG, Brazil
publishDate 2023
dc.date.accessioned.fl_str_mv 2023-08-11T18:49:39Z
dc.date.available.fl_str_mv 2023-08-11T18:49:39Z
dc.date.issued.fl_str_mv 2023
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.citation.fl_str_mv COSTA, Gabriel Luiz et al. Malaria mitochondrial diagnosis: Challenges and Pitfalls. Frontiers in Tropical Diseases, v. 4, p. 1204195, doi.org/10.3389/fitd.2023.1204195.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/60050
dc.identifier.issn.en_US.fl_str_mv 2673-7515
identifier_str_mv COSTA, Gabriel Luiz et al. Malaria mitochondrial diagnosis: Challenges and Pitfalls. Frontiers in Tropical Diseases, v. 4, p. 1204195, doi.org/10.3389/fitd.2023.1204195.
2673-7515
url https://www.arca.fiocruz.br/handle/icict/60050
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Frontiers Media S.A.
publisher.none.fl_str_mv Frontiers Media S.A.
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/60050/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/60050/2/Malaria%20mitochondrial%20diagnosis%20challenges%20and%20pitfalls%20-%20fitd-04-1204195.pdf
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