Development of real-time and lateral flow recombinase polymerase amplification assays for rapid detection of Schistosoma mansoni

Detalhes bibliográficos
Autor(a) principal: Mesquita, Silvia Gonçalves
Data de Publicação: 2022
Outros Autores: Lugli, Elena Birgitta, Matera, Giovanni, Fonseca, Cristina Toscano, Caldeira, Roberta Lima, Webster, Bonnie
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/56536
Resumo: Fundação Oswaldo Cruz. Instituto René Rachou. Grupo de Pesquisa em Helmintologia e Malacologia Médica. Belo Horizonte, MG, Brazil/Wolfson Wellcome Laboratories. Department of Science. Natural History Museum. London, United Kingdom.
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spelling Mesquita, Silvia GonçalvesLugli, Elena BirgittaMatera, GiovanniFonseca, Cristina ToscanoCaldeira, Roberta LimaWebster, Bonnie2023-01-20T16:48:55Z2023-01-20T16:48:55Z2022MESQUITA, Silvia Gonçalves et al. Development of real-time and lateral flow recombinase polymerase amplification assays for rapid detection of Schistosoma mansoni. Front Microbiol., v. 13, 1043596, 2022. doi: 10.3389/fmicb.2022.1043596.1664-302Xhttps://www.arca.fiocruz.br/handle/icict/56536engFrontiers Research FoundationDevelopment of real-time and lateral flow recombinase polymerase amplification assays for rapid detection of Schistosoma mansoniinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto René Rachou. Grupo de Pesquisa em Helmintologia e Malacologia Médica. Belo Horizonte, MG, Brazil/Wolfson Wellcome Laboratories. Department of Science. Natural History Museum. London, United Kingdom.Wolfson Wellcome Laboratories. Department of Science. Natural History Museum. London, United Kingdom.Department of Health Sciences. Unit of Microbiology. University "Magna Graecia" of Catanzaro. Catanzaro, Italy.Fundação Oswaldo Cruz. Instituto René Rachou. Grupo de Pesquisa em Biologia e Imunologia Parasitária. Belo Horizonte, MG, Brazil.Fundação Oswaldo Cruz. Instituto René Rachou. Grupo de Pesquisa em Helmintologia e Malacologia Médica. Belo Horizonte, MG, Brazil.Wolfson Wellcome Laboratories. Department of Science. Natural History Museum. London, United Kingdom.Background: Accurate diagnosis followed by timely treatment is an effective strategy for the prevention of complications together with reducing schistosomiasis transmission. Recombinase Polymerase Amplification (RPA) is a simple, rapid, sensitive, and specific isothermal method with low resource needs. This research aimed at the development and optimisation of a real-time (RT) and a lateral flow (LF) RPA assay for the detection of Schistosoma mansoni. Methodology: Recombinase Polymerase Amplification reactions were performed at full- (as recommended) and half-volumes (to reduce costs), with RT or LF detection systems targeting the S. mansoni mitochondrial minisatellite region. The specificity was assessed using gDNA from other Schistosoma species, helminths co-endemic with S. mansoni, human stool, and urine, and Biomphalaria snail hosts. The analytical sensitivity was evaluated using serial dilutions of gDNA, synthetic copies of the target, and single eggs. The ability of both assays to detect the S. mansoni DNA in human urine and stool samples was also tested. The long-term stability of the RT-RPA reagents was evaluated by storing the reaction components in different temperature conditions for up to 3 weeks. Results: The RT- and the LF-RPA (SmMIT- and SmMIT-LF-RPA, respectively) presented similar results when used full- and half-volumes, thus the latter was followed in all experiments. The SmMIT-RPA was 100% specific to S. mansoni, able to detect a single egg, with a limit of detection (LOD) of down to 1 fg of gDNA and one synthetic copy of the target. The assay was able to detect S. mansoni DNA from stool containing 1 egg/g and in spiked urine at a concentration of 10 fg/μl. SmMIT-RPA reagents were stable for up to 3 weeks when kept at 19°C, and 2 weeks when stored at 27°C. The SmMIT-LF-RPA cross-reacted with Clinostomidae, presented the LOD of 10 fg and one synthetic copy of the target, being able to detect a single egg and 1 egg/g in a stool sample. The LOD in spiked urine samples was 10 pg/μl. Conclusion: The half-volume SmMIT-RPA is a promising method to be used in the field. It is specific, sensitive, robust, and tolerant to inhibitors, with a long-term stability of the reaction components and the real-time visualisation of results.Recombinase Polymerase AmplificationSchistosoma mansoniSchistosomiasisisothermal molecular diagnosticslateral flow RPAmitochondrial minisatellite regionreal-time RPAinfo: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/56536/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALDevelopment of real-time and lateral flow.pdfDevelopment of real-time and lateral flow.pdfapplication/pdf4623810https://www.arca.fiocruz.br/bitstream/icict/56536/2/Development%20of%20real-time%20and%20lateral%20flow.pdfffdd82c29571aeb09d146754ca91dec4MD52icict/565362023-01-20 13:48:56.025oai:www.arca.fiocruz.br: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ório 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dc.title.en_US.fl_str_mv Development of real-time and lateral flow recombinase polymerase amplification assays for rapid detection of Schistosoma mansoni
title Development of real-time and lateral flow recombinase polymerase amplification assays for rapid detection of Schistosoma mansoni
spellingShingle Development of real-time and lateral flow recombinase polymerase amplification assays for rapid detection of Schistosoma mansoni
Mesquita, Silvia Gonçalves
Recombinase Polymerase Amplification
Schistosoma mansoni
Schistosomiasis
isothermal molecular diagnostics
lateral flow RPA
mitochondrial minisatellite region
real-time RPA
title_short Development of real-time and lateral flow recombinase polymerase amplification assays for rapid detection of Schistosoma mansoni
title_full Development of real-time and lateral flow recombinase polymerase amplification assays for rapid detection of Schistosoma mansoni
title_fullStr Development of real-time and lateral flow recombinase polymerase amplification assays for rapid detection of Schistosoma mansoni
title_full_unstemmed Development of real-time and lateral flow recombinase polymerase amplification assays for rapid detection of Schistosoma mansoni
title_sort Development of real-time and lateral flow recombinase polymerase amplification assays for rapid detection of Schistosoma mansoni
author Mesquita, Silvia Gonçalves
author_facet Mesquita, Silvia Gonçalves
Lugli, Elena Birgitta
Matera, Giovanni
Fonseca, Cristina Toscano
Caldeira, Roberta Lima
Webster, Bonnie
author_role author
author2 Lugli, Elena Birgitta
Matera, Giovanni
Fonseca, Cristina Toscano
Caldeira, Roberta Lima
Webster, Bonnie
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Mesquita, Silvia Gonçalves
Lugli, Elena Birgitta
Matera, Giovanni
Fonseca, Cristina Toscano
Caldeira, Roberta Lima
Webster, Bonnie
dc.subject.en.en_US.fl_str_mv Recombinase Polymerase Amplification
Schistosoma mansoni
Schistosomiasis
isothermal molecular diagnostics
lateral flow RPA
mitochondrial minisatellite region
real-time RPA
topic Recombinase Polymerase Amplification
Schistosoma mansoni
Schistosomiasis
isothermal molecular diagnostics
lateral flow RPA
mitochondrial minisatellite region
real-time RPA
description Fundação Oswaldo Cruz. Instituto René Rachou. Grupo de Pesquisa em Helmintologia e Malacologia Médica. Belo Horizonte, MG, Brazil/Wolfson Wellcome Laboratories. Department of Science. Natural History Museum. London, United Kingdom.
publishDate 2022
dc.date.issued.fl_str_mv 2022
dc.date.accessioned.fl_str_mv 2023-01-20T16:48:55Z
dc.date.available.fl_str_mv 2023-01-20T16:48:55Z
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 MESQUITA, Silvia Gonçalves et al. Development of real-time and lateral flow recombinase polymerase amplification assays for rapid detection of Schistosoma mansoni. Front Microbiol., v. 13, 1043596, 2022. doi: 10.3389/fmicb.2022.1043596.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/56536
dc.identifier.issn.en_US.fl_str_mv 1664-302X
identifier_str_mv MESQUITA, Silvia Gonçalves et al. Development of real-time and lateral flow recombinase polymerase amplification assays for rapid detection of Schistosoma mansoni. Front Microbiol., v. 13, 1043596, 2022. doi: 10.3389/fmicb.2022.1043596.
1664-302X
url https://www.arca.fiocruz.br/handle/icict/56536
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/56536/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/56536/2/Development%20of%20real-time%20and%20lateral%20flow.pdf
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repository.name.fl_str_mv Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)
repository.mail.fl_str_mv repositorio.arca@fiocruz.br
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