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AutorAmaral, Lara Cotta
AutorRobortella, Daniela Rocha
AutorGuimarães, Luiz Felipe Ferreira
AutorLimongi, Jean Ezequiel
AutorFontes, Cor Jesus Fernandes
AutorPereira, Dhelio Batista
AutorBrito, Cristiana Ferreira Alves de
AutorKano, Flora Satiko
AutorSousa, Taís Nóbrega de
AutorCarvalho, Luzia Helena de
Fecha de acceso2019-10-07T19:22:15Z
Fecha de disponibilización2019-10-07T19:22:15Z
Fecha de publicación2019
ReferenciaAMARAL, Lara Cotta et al. Ribosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infections. Malaria Journal, v. 18, n. 1, p. 1-14, 2019.pt_BR
ISSN1475-2875pt_BR
URIhttps://www.arca.fiocruz.br/handle/icict/36232
Idiomaengpt_BR
EditorBMCpt_BR
Derechos de autorrestricted accesspt_BR
Palabras clave en PortuguésMalariapt_BR
Palabras clave en PortuguésPCRpt_BR
TítuloRibosomal and non-ribosomal PCR targets for the detection of low-density and mixed malaria infectionspt_BR
Tipo del documentoArticle
DOI10.1186/s12936-019-2781-3
Resumen en InglésBackground The 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 targets—ribosomal (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. Methods A 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. Results The 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 (~ 20%) identified only by the NR-qPCR. Conclusions This 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. Electronic supplementary material The online version of this article (10.1186/s12936-019-2781-3) contains supplementary material, which is available to authorized users.pt_BR
AfiliaciónFundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil/Universidade Federal de Minas Gerais. Belo Horizonte, MG, Brasil.pt_BR
AfiliaciónFundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil/Universidade Federal de Minas Gerais. Belo Horizonte, MG, Brasil.pt_BR
AfiliaciónFundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil.pt_BR
AfiliaciónUniversidade Federal de Uberlândia. Uberlândia, MG, Brasil.pt_BR
AfiliaciónUniversidade Federal de Mato Grosso.Hospital Julio Mueller. Cuiabá. MT, Brasil.pt_BR
AfiliaciónCentro de Pesquisas em Medicina Tropical de Rondônia. Porto Velho, RO, Brasil.pt_BR
AfiliaciónFundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil.pt_BR
AfiliaciónFundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil.pt_BR
AfiliaciónFundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil.pt_BR
AfiliaciónFundação Oswaldo Cruz. Instituto Rene Rachou. Belo Horizonte, MG, Brasil / Universidade Federal de Minas Gerais. Belo Horizonte, MG, Brasil.pt_BR
Palavras clave en InglêsMalariapt_BR
Palavras clave en InglêsMolecular diagnosispt_BR
Palavras clave en InglêsPCRpt_BR
Palavras clave en InglêsSubmicroscopicpt_BR
Palavras clave en InglêsMixed-malaria infectionspt_BR
Fecha de embargo2022-01-01


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