Development and Evaluation of a Sensitive PCR-ELISA System for Detection of Schistosoma Infection in Feces

Detalhes bibliográficos
Autor(a) principal: Gomes, Luciana Inácia
Data de Publicação: 2010
Outros Autores: Marques, Letícia Helena dos Santos, Enk, Martin Johannes, Oliveira, Maria Cláudia de, Coelho, Paulo Marcos Zach, Rabello, Ana Lúcia Teles
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/5622
Resumo: Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Pesquisas Clínicas. Belo Horizonte, MG, Brazil
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spelling Gomes, Luciana InáciaMarques, Letícia Helena dos SantosEnk, Martin JohannesOliveira, Maria Cláudia deCoelho, Paulo Marcos ZachRabello, Ana Lúcia Teles2012-10-01T17:16:47Z2012-10-01T17:16:47Z2010GOMES, Luciana Inácia et al. Development and Evaluation of a Sensitive PCR-ELISA System for Detection of Schistosoma Infection in Feces.PLoS Negl Trop Dis. 2010 Apr 20;4(4):e664.19352727https://www.arca.fiocruz.br/handle/icict/562210.1371/journal.pntd.0000664engPublic Library of ScienceDevelopment and Evaluation of a Sensitive PCR-ELISA System for Detection of Schistosoma Infection in Fecesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Pesquisas Clínicas. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Pesquisas Clínicas. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Esquistossomose. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Pesquisas Clínicas. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Esquistossomose. Belo Horizonte, MG, BrazilFundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Pesquisas Clínicas. Belo Horizonte, MG, BrazilBackground: A PCR-enzyme-linked immunosorbent assay (PCR-ELISA) was developed to overcome the need for sensitive techniques for the efficient diagnosis of Schistosoma infection in endemic settings with low parasitic burden. Methodology/Principal Findings: This system amplifies a 121-base pair tandem repeat DNA sequence, immobilizes the resultant 59 biotinylated product on streptavidin-coated strip-well microplates and uses anti-fluorescein antibodies conjugated to horseradish peroxidase to detect the hybridized fluorescein-labeled oligonucleotide probe. The detection limit of the Schistosoma PCR-ELISA system was determined to be 1.3 fg of S. mansoni genomic DNA (less than the amount found in a single cell) and estimated to be 0.15 S. mansoni eggs per gram of feces (fractions of an egg). The system showed good precision and genus specificity since the DNA target was found in seven Schistosoma DNA samples: S. mansoni, S. haematobium, S. bovis, S. intercalatum, S. japonicum, S. magrebowiei and S. rhodaini. By evaluating 206 patients living in an endemic area in Brazil, the prevalence of S. mansoni infection was determined to be 18% by examining 12 Kato-Katz slides (41.7 mg/smear, 500 mg total) of a single fecal sample from each person, while the Schistosoma PCR-ELISA identified a 30% rate of infection using 500-mg of the same fecal sample. When considering the Kato-Katz method as the reference test, artificial sensitivity and specificity rates of the PCR-ELISA system were 97.4% and 85.1%, respectively. The potential for estimating parasitic load by DNA detection in feces was assessed by comparing absorbance values and eggs per gram of feces, with a Spearman correlation coefficient of 0.700 (P,0.0001). Conclusions/Significance: This study reports the development and field evaluation of a sensitive Schistosoma PCR-ELISA, a system that may serve as an alternative for diagnosing Schistosoma infection.Neglected diseasePCR-ELISASchistosoma DNAinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZORIGINAL52.2010.pdf52.2010.pdfapplication/pdf910859https://www.arca.fiocruz.br/bitstream/icict/5622/1/52.2010.pdfeca5b99ef29c8b9bdc7bcf284f8c2d93MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-82008https://www.arca.fiocruz.br/bitstream/icict/5622/2/license.txt06db3f0df647fd9ddff657403bbe0ee2MD52TEXT52.2010.pdf.txt52.2010.pdf.txtExtracted texttext/plain46700https://www.arca.fiocruz.br/bitstream/icict/5622/5/52.2010.pdf.txt011b9908a6d05c378bc253646b1b9b0eMD55THUMBNAIL52.2010.pdf.jpg52.2010.pdf.jpgGenerated Thumbnailimage/jpeg2600https://www.arca.fiocruz.br/bitstream/icict/5622/4/52.2010.pdf.jpgd210189baf8c9f7ed526ad5fd48a5e53MD54icict/56222018-05-16 10:34:55.681oai:www.arca.fiocruz.br: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Repositório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352018-05-16T13:34:55Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)false
dc.title.pt_BR.fl_str_mv Development and Evaluation of a Sensitive PCR-ELISA System for Detection of Schistosoma Infection in Feces
title Development and Evaluation of a Sensitive PCR-ELISA System for Detection of Schistosoma Infection in Feces
spellingShingle Development and Evaluation of a Sensitive PCR-ELISA System for Detection of Schistosoma Infection in Feces
Gomes, Luciana Inácia
Neglected disease
PCR-ELISA
Schistosoma DNA
title_short Development and Evaluation of a Sensitive PCR-ELISA System for Detection of Schistosoma Infection in Feces
title_full Development and Evaluation of a Sensitive PCR-ELISA System for Detection of Schistosoma Infection in Feces
title_fullStr Development and Evaluation of a Sensitive PCR-ELISA System for Detection of Schistosoma Infection in Feces
title_full_unstemmed Development and Evaluation of a Sensitive PCR-ELISA System for Detection of Schistosoma Infection in Feces
title_sort Development and Evaluation of a Sensitive PCR-ELISA System for Detection of Schistosoma Infection in Feces
author Gomes, Luciana Inácia
author_facet Gomes, Luciana Inácia
Marques, Letícia Helena dos Santos
Enk, Martin Johannes
Oliveira, Maria Cláudia de
Coelho, Paulo Marcos Zach
Rabello, Ana Lúcia Teles
author_role author
author2 Marques, Letícia Helena dos Santos
Enk, Martin Johannes
Oliveira, Maria Cláudia de
Coelho, Paulo Marcos Zach
Rabello, Ana Lúcia Teles
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Gomes, Luciana Inácia
Marques, Letícia Helena dos Santos
Enk, Martin Johannes
Oliveira, Maria Cláudia de
Coelho, Paulo Marcos Zach
Rabello, Ana Lúcia Teles
dc.subject.en.pt_BR.fl_str_mv Neglected disease
PCR-ELISA
Schistosoma DNA
topic Neglected disease
PCR-ELISA
Schistosoma DNA
description Fundação Oswaldo Cruz. Centro de Pesquisas René Rachou. Laboratório de Pesquisas Clínicas. Belo Horizonte, MG, Brazil
publishDate 2010
dc.date.issued.fl_str_mv 2010
dc.date.accessioned.fl_str_mv 2012-10-01T17:16:47Z
dc.date.available.fl_str_mv 2012-10-01T17:16:47Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
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dc.identifier.citation.fl_str_mv GOMES, Luciana Inácia et al. Development and Evaluation of a Sensitive PCR-ELISA System for Detection of Schistosoma Infection in Feces.PLoS Negl Trop Dis. 2010 Apr 20;4(4):e664.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/5622
dc.identifier.issn.none.fl_str_mv 19352727
dc.identifier.doi.none.fl_str_mv 10.1371/journal.pntd.0000664
identifier_str_mv GOMES, Luciana Inácia et al. Development and Evaluation of a Sensitive PCR-ELISA System for Detection of Schistosoma Infection in Feces.PLoS Negl Trop Dis. 2010 Apr 20;4(4):e664.
19352727
10.1371/journal.pntd.0000664
url https://www.arca.fiocruz.br/handle/icict/5622
dc.language.iso.fl_str_mv eng
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dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
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