Ribosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infections

Detalhes bibliográficos
Autor(a) principal: Amaral, Lara Cotta
Data de Publicação: 2019
Outros Autores: Robortella, Daniela Rocha, Guimarães, Luiz Felipe Ferreira, Limongi, Jean Ezequiel, Fontes, Cor Jesus Fernandes, Pereira, Dhelio Batista, Kano, Flora Satiko, Sousa, Taís Nóbrega de, Carvalho, Luzia Helena
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/55148
Resumo: Fundação Oswaldo Cruz. Instituto René Rachou. Belo Horizonte, MG, Brazil
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spelling Amaral, Lara CottaRobortella, Daniela RochaGuimarães, Luiz Felipe FerreiraLimongi, Jean EzequielFontes, Cor Jesus FernandesPereira, Dhelio BatistaPereira, Dhelio BatistaKano, Flora SatikoSousa, Taís Nóbrega deCarvalho, Luzia Helena2022-10-14T16:58:37Z2022-10-14T16:58:37Z2019AMARAL, Lara Cota et al. Ribosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infections. Malar J, v. 18, n. 1, 154, 2019. doi.org/10.1186/s12936-019-2781-31475-2875https://www.arca.fiocruz.br/handle/icict/5514810.1186/s12936-019-2781-3engBioMed CentralRibosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infectionsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto René Rachou. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Instituto René Rachou. Belo Horizonte, MG, Brazil/Universidade Federal de Minas Gerais. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Instituto René Rachou. Belo Horizonte, MG, BrazilUniversidade Federal de Uberlândia. Uberlândia, MG, BrazilUniversidade Federal do Mato Grosso. Hospital Júlio Müller. Cuiabá, MT, BrazilCentro de Pesquisas em Medicina Tropical de Rondônia. Porto Velho, RO, BrazilCentro de Pesquisas em Medicina Tropical de Rondônia. Porto Velho, RO, BrazilFundação Oswaldo Cruz. Instituto René Rachou. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Instituto René Rachou. Belo Horizonte, MG, Brazil/Universidade Federal de Minas Gerais. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Instituto René Rachou. Belo Horizonte, MG, BrazilBackgroundThe unexpected high proportion of submicroscopic malaria infections in areas with low transmission intensity challenges the control and elimination of malaria in the Americas. The current PCR-based assays present limitations as most protocols still rely on amplification of few-copies target gene. Here, the hypothesis was that amplification of different plasmodial targetsribosomal (18S rRNA) and non-ribosomal multi-copy sequences (Pvr47 for Plasmodium vivax and Pfr364 for Plasmodium falciparum)could increase the chances of detecting submicroscopic malaria infection.MethodsA non-ribosomal real-time PCR assay targeting Pvr47/Pfr364 (NR-qPCR) was established and compared with three additional PCR protocols, two of them based on 18S rRNA gene amplification (Nested-PCR and R-qPCR) and one based on Pvr47/Pfr364 targets (NR-cPCR). The limit of detection of each PCR protocol, at single and artificial mixed P. vivax/P. falciparum infections, was determined by end-point titration curves. Field samples from clinical (n=110) and subclinical (n=324) malaria infections were used to evaluate the impact of using multiple molecular targets to detect malaria infections.ResultsThe results demonstrated that an association of ribosomal and non-ribosomal targets did not increase sensitivity to detect submicroscopic malaria infections. Despite of that, artificial mixed-malaria infections demonstrated that the NR-qPCR was the most sensitive protocol to detect low-levels of P. vivax/P. falciparum co-infections. Field studies confirmed that submicroscopic malaria represented a large proportion (up to 77%) of infections among asymptomatic Amazonian residents, with a high proportion of infections (similar to 20%) identified only by the NR-qPCR.ConclusionsThis study presents a new species-specific non-ribosomal PCR assay with potential to identify low-density P. vivax and P. falciparum infections. As the majority of subclinical infections was caused by P. vivax, the commonest form of malaria in the Amazon area, future studies should investigate the potential of Pvr47/Pfr364 to detect mixed-malaria infections in the field.MalariaMolecular diagnosisPCRSubmicroscopicMixed-malaria infectionsinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZORIGINALRibosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infections.pdfRibosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infections.pdfapplication/pdf1373296https://www.arca.fiocruz.br/bitstream/icict/55148/2/Ribosomal%20and%20non-ribosomal%20PCR%20targets%20for%20the%20detection%20of%20low-density%20and%20mixed%20malaria%20infections.pdf73e6cd514d4d18db48458f202a33a442MD52LICENSElicense.txttext/plain1748https://www.arca.fiocruz.br/bitstream/icict/55148/1/license.txt8a4605be74aa9ea9d79846c1fba20a33MD51icict/551482022-10-14 13:58:38.481oai:www.arca.fiocruz.br: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Repositório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352022-10-14T16:58:38Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)false
dc.title.en_US.fl_str_mv Ribosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infections
title Ribosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infections
spellingShingle Ribosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infections
Amaral, Lara Cotta
Malaria
Molecular diagnosis
PCR
Submicroscopic
Mixed-malaria infections
title_short Ribosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infections
title_full Ribosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infections
title_fullStr Ribosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infections
title_full_unstemmed Ribosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infections
title_sort Ribosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infections
author Amaral, Lara Cotta
author_facet Amaral, Lara Cotta
Robortella, Daniela Rocha
Guimarães, Luiz Felipe Ferreira
Limongi, Jean Ezequiel
Fontes, Cor Jesus Fernandes
Pereira, Dhelio Batista
Kano, Flora Satiko
Sousa, Taís Nóbrega de
Carvalho, Luzia Helena
author_role author
author2 Robortella, Daniela Rocha
Guimarães, Luiz Felipe Ferreira
Limongi, Jean Ezequiel
Fontes, Cor Jesus Fernandes
Pereira, Dhelio Batista
Kano, Flora Satiko
Sousa, Taís Nóbrega de
Carvalho, Luzia Helena
author2_role author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Amaral, Lara Cotta
Robortella, Daniela Rocha
Guimarães, Luiz Felipe Ferreira
Limongi, Jean Ezequiel
Fontes, Cor Jesus Fernandes
Pereira, Dhelio Batista
Pereira, Dhelio Batista
Kano, Flora Satiko
Sousa, Taís Nóbrega de
Carvalho, Luzia Helena
dc.subject.en.en_US.fl_str_mv Malaria
Molecular diagnosis
PCR
Submicroscopic
Mixed-malaria infections
topic Malaria
Molecular diagnosis
PCR
Submicroscopic
Mixed-malaria infections
description Fundação Oswaldo Cruz. Instituto René Rachou. Belo Horizonte, MG, Brazil
publishDate 2019
dc.date.issued.fl_str_mv 2019
dc.date.accessioned.fl_str_mv 2022-10-14T16:58:37Z
dc.date.available.fl_str_mv 2022-10-14T16:58:37Z
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 AMARAL, Lara Cota et al. Ribosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infections. Malar J, v. 18, n. 1, 154, 2019. doi.org/10.1186/s12936-019-2781-3
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/55148
dc.identifier.issn.en_US.fl_str_mv 1475-2875
dc.identifier.doi.none.fl_str_mv 10.1186/s12936-019-2781-3
identifier_str_mv AMARAL, Lara Cota et al. Ribosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infections. Malar J, v. 18, n. 1, 154, 2019. doi.org/10.1186/s12936-019-2781-3
1475-2875
10.1186/s12936-019-2781-3
url https://www.arca.fiocruz.br/handle/icict/55148
dc.language.iso.fl_str_mv eng
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dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv BioMed Central
publisher.none.fl_str_mv BioMed Central
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
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instname_str Fundação Oswaldo Cruz (FIOCRUZ)
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bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/55148/2/Ribosomal%20and%20non-ribosomal%20PCR%20targets%20for%20the%20detection%20of%20low-density%20and%20mixed%20malaria%20infections.pdf
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