Malaria mitochondrial diagnosis: Challenges and Pitfalls.
Autor(a) principal: | |
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Data de Publicação: | 2023 |
Outros Autores: | , , , , , , , , , , , , |
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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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 |
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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 |
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eng |
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eng |
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info:eu-repo/semantics/openAccess |
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openAccess |
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Frontiers Media S.A. |
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Frontiers Media S.A. |
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reponame:Repositório Institucional da FIOCRUZ (ARCA) instname:Fundação Oswaldo Cruz (FIOCRUZ) instacron:FIOCRUZ |
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FIOCRUZ |
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