Improving the Molecular Diagnosis of Malaria: Droplet Digital PCR-Based Method Using Saliva as a DNA Source

Detalhes bibliográficos
Autor(a) principal: Costa, Gabriel Luíz
Data de Publicação: 2022
Outros Autores: Alvarenga, Denise Anete Madureira, Aguiar, Anna Caroline Campos, Louzada, Jaime, Pereira, Dhélio Batista, Oliveira, Tatiana Flávia de, Fonseca Júnior, Antônio Augusto, Carvalho, Luzia Helena, Brito, Cristiana Ferreira Alves de, Sousa, Taís Nóbrega de
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/55684
Resumo: Fundação Oswaldo Cruz. Instituto René Rachou. Molecular Biology and Malaria Immunology Research Group. Belo Horizonte, MG, Brazil.
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spelling Costa, Gabriel LuízAlvarenga, Denise Anete MadureiraAguiar, Anna Caroline CamposLouzada, JaimePereira, Dhélio BatistaOliveira, Tatiana Flávia deFonseca Júnior, Antônio AugustoCarvalho, Luzia HelenaBrito, Cristiana Ferreira Alves deSousa, Taís Nóbrega de2022-11-17T17:52:24Z2022-11-17T17:52:24Z2022COSTA, Gabriel Luíz et al. Improving the Molecular Diagnosis of Malaria: Droplet Digital PCR-Based Method Using Saliva as a DNA Source. Frontiers in Microbiology, v. 13, 882530, p. 1-12, May 2022.1664-302Xhttps://www.arca.fiocruz.br/handle/icict/5568410.3389/fmicb.2022.882530engFrontiers Research FoundationImproving the Molecular Diagnosis of Malaria: Droplet Digital PCR-Based Method Using Saliva as a DNA Sourceinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto René Rachou. Molecular Biology and Malaria Immunology Research Group. Belo Horizonte, MG, Brazil.Fundação Oswaldo Cruz. Instituto René Rachou. Molecular Biology and Malaria Immunology Research Group. Belo Horizonte, MG, Brazil.University of São Paulo. Department of Bioscience. Federal. Santos, SP, Brazil.Federal University of Roraima. Health Sciences Center. Boa Vista, RO, Brazil.Center for Research in Tropical Medicine. Porto Velho, RO, Brazil.Laboratório Federal de Defesa Agropecuária de Minas Gerais. Pedro Leopoldo, MG, Brazil.Laboratório Federal de Defesa Agropecuária de Minas Gerais. Pedro Leopoldo, MG, BrazilFundação Oswaldo Cruz. Instituto René Rachou. Molecular Biology and Malaria Immunology Research Group. Belo Horizonte, MG, Brazil.Fundação Oswaldo Cruz. Instituto René Rachou. Molecular Biology and Malaria Immunology Research Group. Belo Horizonte, MG, Brazil.Fundação Oswaldo Cruz. Instituto René Rachou. Molecular Biology and Malaria Immunology Research Group. Belo Horizonte, MG, Brazil.Malaria is an acute febrile disease caused by a protozoan of the genus Plasmodium. Light microscopy (LM) is the gold standard for the diagnosis of malaria. Despite this method being rapid and inexpensive, it has a low limit of detection, which hampers the identification of low parasitemia infections. By using multicopy targets and highly sensitive molecular techniques, it is possible to change this scenario. In this study, we evaluated the performance of droplet digital PCR (ddPCR) to detect Plasmodium DNA obtained from saliva samples (whole saliva and buccal swab) of 157 individuals exposed to malaria transmission from the Brazilian Amazon region. We used the highly sensitive ddPCR method with non-ribosomal multicopy targets for Plasmodium vivax (Pvr47) and Plasmodium falciparum (Pfr364). There was good concordance between the quantitative real-time PCR (qPCR) results from the saliva and blood, except for mixed-species infections. The sensitivity of qPCR was 93% for blood, 77% for saliva, and 47% for swabs. Parasite DNA was not detected in saliva samples in low-density infections compared with the detection in blood samples. ddPCR showed increased sensitivity for detecting Plasmodium in the blood and swabs (99% in blood, 73% in saliva, and 59% in swabs). Notably, ddPCR detected more mixed infections in the blood (15%), saliva (9%), and swabs (18%) than qPCR. Our data showed that the differences between ddPCR and qPCR were the result of a higher number of P. falciparum infections detected by ddPCR. Overall, there was a moderate correlation between parasite densities estimated by the different methods in the blood. Our findings highlight the possibility of using non-invasive sample collection methods for malaria diagnosis by targeting multicopy sequences combined with highly sensitive molecular methods.malariaMolecular diagnosisDroplet digital PCRSalivaPlasmodium falciparumPlasmodium vivaxinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZORIGINALImproving the Molecular Diagnosis of Malaria.pdfImproving the Molecular Diagnosis of Malaria.pdfapplication/pdf1646316https://www.arca.fiocruz.br/bitstream/icict/55684/2/Improving%20the%20Molecular%20Diagnosis%20of%20Malaria.pdfe33f000e337665312d09a0b25c5d995dMD52LICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/55684/1/license.txt5a560609d32a3863062d77ff32785d58MD51icict/556842022-11-18 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dc.title.en_US.fl_str_mv Improving the Molecular Diagnosis of Malaria: Droplet Digital PCR-Based Method Using Saliva as a DNA Source
title Improving the Molecular Diagnosis of Malaria: Droplet Digital PCR-Based Method Using Saliva as a DNA Source
spellingShingle Improving the Molecular Diagnosis of Malaria: Droplet Digital PCR-Based Method Using Saliva as a DNA Source
Costa, Gabriel Luíz
malaria
Molecular diagnosis
Droplet digital PCR
Saliva
Plasmodium falciparum
Plasmodium vivax
title_short Improving the Molecular Diagnosis of Malaria: Droplet Digital PCR-Based Method Using Saliva as a DNA Source
title_full Improving the Molecular Diagnosis of Malaria: Droplet Digital PCR-Based Method Using Saliva as a DNA Source
title_fullStr Improving the Molecular Diagnosis of Malaria: Droplet Digital PCR-Based Method Using Saliva as a DNA Source
title_full_unstemmed Improving the Molecular Diagnosis of Malaria: Droplet Digital PCR-Based Method Using Saliva as a DNA Source
title_sort Improving the Molecular Diagnosis of Malaria: Droplet Digital PCR-Based Method Using Saliva as a DNA Source
author Costa, Gabriel Luíz
author_facet Costa, Gabriel Luíz
Alvarenga, Denise Anete Madureira
Aguiar, Anna Caroline Campos
Louzada, Jaime
Pereira, Dhélio Batista
Oliveira, Tatiana Flávia de
Fonseca Júnior, Antônio Augusto
Carvalho, Luzia Helena
Brito, Cristiana Ferreira Alves de
Sousa, Taís Nóbrega de
author_role author
author2 Alvarenga, Denise Anete Madureira
Aguiar, Anna Caroline Campos
Louzada, Jaime
Pereira, Dhélio Batista
Oliveira, Tatiana Flávia de
Fonseca Júnior, Antônio Augusto
Carvalho, Luzia Helena
Brito, Cristiana Ferreira Alves de
Sousa, Taís Nóbrega de
author2_role author
author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Costa, Gabriel Luíz
Alvarenga, Denise Anete Madureira
Aguiar, Anna Caroline Campos
Louzada, Jaime
Pereira, Dhélio Batista
Oliveira, Tatiana Flávia de
Fonseca Júnior, Antônio Augusto
Carvalho, Luzia Helena
Brito, Cristiana Ferreira Alves de
Sousa, Taís Nóbrega de
dc.subject.en.en_US.fl_str_mv malaria
Molecular diagnosis
Droplet digital PCR
Saliva
Plasmodium falciparum
Plasmodium vivax
topic malaria
Molecular diagnosis
Droplet digital PCR
Saliva
Plasmodium falciparum
Plasmodium vivax
description Fundação Oswaldo Cruz. Instituto René Rachou. Molecular Biology and Malaria Immunology Research Group. Belo Horizonte, MG, Brazil.
publishDate 2022
dc.date.accessioned.fl_str_mv 2022-11-17T17:52:24Z
dc.date.available.fl_str_mv 2022-11-17T17:52:24Z
dc.date.issued.fl_str_mv 2022
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 Luíz et al. Improving the Molecular Diagnosis of Malaria: Droplet Digital PCR-Based Method Using Saliva as a DNA Source. Frontiers in Microbiology, v. 13, 882530, p. 1-12, May 2022.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/55684
dc.identifier.issn.en_US.fl_str_mv 1664-302X
dc.identifier.doi.none.fl_str_mv 10.3389/fmicb.2022.882530
identifier_str_mv COSTA, Gabriel Luíz et al. Improving the Molecular Diagnosis of Malaria: Droplet Digital PCR-Based Method Using Saliva as a DNA Source. Frontiers in Microbiology, v. 13, 882530, p. 1-12, May 2022.
1664-302X
10.3389/fmicb.2022.882530
url https://www.arca.fiocruz.br/handle/icict/55684
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 Research Foundation
publisher.none.fl_str_mv Frontiers Research Foundation
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/55684/2/Improving%20the%20Molecular%20Diagnosis%20of%20Malaria.pdf
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repository.name.fl_str_mv Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)
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