Analytical validation of real-time quantitative PCR assays for optimum diagnosis of vivax malaria

Detalhes bibliográficos
Autor(a) principal: Oliveira, Natália Ketrin Almeida de
Data de Publicação: 2019
Outros Autores: Moreira, Otacilio C., Lavigne, Aline Rosa de, Lima, Leila Mendonça, Werneck, Guilherme Loureiro, Daniel-Ribeiro, Cláudio Tadeu, Cruz, Maria de Fátima Ferreira da
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/34991
Resumo: Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Malária. Rio de Janeiro, RJ, Brasil.
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spelling Oliveira, Natália Ketrin Almeida deMoreira, Otacilio C.Lavigne, Aline Rosa deLima, Leila MendonçaWerneck, Guilherme LoureiroDaniel-Ribeiro, Cláudio TadeuCruz, Maria de Fátima Ferreira da2019-08-22T17:42:31Z2019-08-22T17:42:31Z2019OLIVEIRA, Natália Ketrin Almeida de et al. Analytical validation of real-time quantitative PCR assays for optimum diagnosis of vivax malaria. Memórias do Instituto Oswaldo Cruz, Rio de Janeiro, v. 114, p. 1-6, 2019.0074-0276https://www.arca.fiocruz.br/handle/icict/3499110.1590/0074-027601803501678-8060engFundação Oswaldo Cruz. Instituto Oswaldo CruzMaláriaDiagnósticoPlasmodium vivaxPlasmodium vivaxMalariaDiagnosisAnalytical validation of real-time quantitative PCR assays for optimum diagnosis of vivax malariainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Malária. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Malária. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Genômica Funcional e Bioinformática. Rio de Janeiro, RJ, Brasil.Universidade do Estado do Rio de Janeiro. Instituto de Medicina Social. Departamento de Epidemiologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Malária. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Malária. Rio de Janeiro, RJ, Brasil.BACKGROUND The prompt diagnosis of plasmodial species for effective treatment prevents worsening of individual health and avoids transmission maintenance or even malaria reintroduction in areas where Plasmodium does not exist. Polymerase chain reaction (PCR) allows for the detection of parasites below the threshold of microscopic examination. OBJECTIVE Our aim was to develop a real-time PCR test to reduce diagnostic errors and increase efficacy. METHODS The lower limit of quantification and the linearity/analytical sensitivity to measure sensitivity or limit of detection (LoD) were determined. Intra-assay variations (repeatability) and alterations between assays, operators, and instruments (reproducibility) were also assessed to set precision. FINDINGS The linearity in SYBR™ Green and TaqMan™ systems was 106 and 102 copies and analytical sensitivity 1.13 and 1.17 copies/μL, respectively. Real-time PCR was more sensitive than conventional PCR, showing a LoD of 0.01 parasite (p)/μL. Reproducibility and repeatability (precision) were 100% for up to 0.1 p/μL in SYBR™ Green and 1 p/μL in TaqMan™ and conventional PCR. CONCLUSION Real-time PCR may replace conventional PCR in reference laboratories for P. vivax detection due to its rapidity. The TaqMan™ system is the most indicated when quantification assays are required. Performing tests in triplicate when diagnosing Plasmodium-infected-asymptomatic individuals is recommended to minimise diagnostic errors.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/34991/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALAlineLavigne_OtacilioMoreira_etal_IOC_2019.pdfAlineLavigne_OtacilioMoreira_etal_IOC_2019.pdfapplication/pdf458878https://www.arca.fiocruz.br/bitstream/icict/34991/2/AlineLavigne_OtacilioMoreira_etal_IOC_2019.pdf4993b198c5212641c504f2cc9791d069MD52TEXTAlineLavigne_OtacilioMoreira_etal_IOC_2019.pdf.txtAlineLavigne_OtacilioMoreira_etal_IOC_2019.pdf.txtExtracted texttext/plain35287https://www.arca.fiocruz.br/bitstream/icict/34991/3/AlineLavigne_OtacilioMoreira_etal_IOC_2019.pdf.txt1635cbeacd5c53768e910b0619f0c9c5MD53icict/349912022-06-24 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dc.title.pt_BR.fl_str_mv Analytical validation of real-time quantitative PCR assays for optimum diagnosis of vivax malaria
title Analytical validation of real-time quantitative PCR assays for optimum diagnosis of vivax malaria
spellingShingle Analytical validation of real-time quantitative PCR assays for optimum diagnosis of vivax malaria
Oliveira, Natália Ketrin Almeida de
Malária
Diagnóstico
Plasmodium vivax
Plasmodium vivax
Malaria
Diagnosis
title_short Analytical validation of real-time quantitative PCR assays for optimum diagnosis of vivax malaria
title_full Analytical validation of real-time quantitative PCR assays for optimum diagnosis of vivax malaria
title_fullStr Analytical validation of real-time quantitative PCR assays for optimum diagnosis of vivax malaria
title_full_unstemmed Analytical validation of real-time quantitative PCR assays for optimum diagnosis of vivax malaria
title_sort Analytical validation of real-time quantitative PCR assays for optimum diagnosis of vivax malaria
author Oliveira, Natália Ketrin Almeida de
author_facet Oliveira, Natália Ketrin Almeida de
Moreira, Otacilio C.
Lavigne, Aline Rosa de
Lima, Leila Mendonça
Werneck, Guilherme Loureiro
Daniel-Ribeiro, Cláudio Tadeu
Cruz, Maria de Fátima Ferreira da
author_role author
author2 Moreira, Otacilio C.
Lavigne, Aline Rosa de
Lima, Leila Mendonça
Werneck, Guilherme Loureiro
Daniel-Ribeiro, Cláudio Tadeu
Cruz, Maria de Fátima Ferreira da
author2_role author
author
author
author
author
author
dc.contributor.author.fl_str_mv Oliveira, Natália Ketrin Almeida de
Moreira, Otacilio C.
Lavigne, Aline Rosa de
Lima, Leila Mendonça
Werneck, Guilherme Loureiro
Daniel-Ribeiro, Cláudio Tadeu
Cruz, Maria de Fátima Ferreira da
dc.subject.other.pt_BR.fl_str_mv Malária
Diagnóstico
Plasmodium vivax
topic Malária
Diagnóstico
Plasmodium vivax
Plasmodium vivax
Malaria
Diagnosis
dc.subject.en.pt_BR.fl_str_mv Plasmodium vivax
Malaria
Diagnosis
description Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisa em Malária. Rio de Janeiro, RJ, Brasil.
publishDate 2019
dc.date.accessioned.fl_str_mv 2019-08-22T17:42:31Z
dc.date.available.fl_str_mv 2019-08-22T17:42:31Z
dc.date.issued.fl_str_mv 2019
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 OLIVEIRA, Natália Ketrin Almeida de et al. Analytical validation of real-time quantitative PCR assays for optimum diagnosis of vivax malaria. Memórias do Instituto Oswaldo Cruz, Rio de Janeiro, v. 114, p. 1-6, 2019.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/34991
dc.identifier.issn.pt_BR.fl_str_mv 0074-0276
dc.identifier.doi.none.fl_str_mv 10.1590/0074-02760180350
dc.identifier.eissn.none.fl_str_mv 1678-8060
identifier_str_mv OLIVEIRA, Natália Ketrin Almeida de et al. Analytical validation of real-time quantitative PCR assays for optimum diagnosis of vivax malaria. Memórias do Instituto Oswaldo Cruz, Rio de Janeiro, v. 114, p. 1-6, 2019.
0074-0276
10.1590/0074-02760180350
1678-8060
url https://www.arca.fiocruz.br/handle/icict/34991
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 Fundação Oswaldo Cruz. Instituto Oswaldo Cruz
publisher.none.fl_str_mv Fundação Oswaldo Cruz. Instituto Oswaldo Cruz
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)
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