Development of a loop-mediated isothermal amplification (LAMP) assay for the detection of Anaplasma marginale

Detalhes bibliográficos
Autor(a) principal: Giglioti, Rodrigo [UNESP]
Data de Publicação: 2019
Outros Autores: Bassetto, Cesar Cristiano [UNESP], Okino, Cintia Hiromi, Oliveira, Henrique Nunes de [UNESP], Sena Oliveira, Marcia Cristina de
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1007/s10493-018-0327-y
http://hdl.handle.net/11449/186631
Resumo: Parasitemia generated by Anaplasma marginale causes significant losses in the cattle industry. A major constraint to the effective control and management of anaplasmosis in livestock is the lack of a rapid and reliable diagnostic test to identify the parasite and allow for immediate therapy. In the present study, we developed a novel DNA-based assay for the detection of A. marginale in bovine blood samples, using loop-mediated isothermal amplification (LAMP). DNA from six cattle and hemoparasite samples (Babesia bovis, Babesia bigemina, Anaplasma centrale and A. marginale) were tested for specificity, sensitivity and cross-reactions. The developed LAMP procedures were also confirmed and compared with the qPCR method. The same gene sequence (major surface protein 1b, msp1b) of A. marginale was used to design a set of primers for the LAMP and qPCR assays. The results showed that LAMP is specific, as no positive signal was observed for the other tested hemoparasites. However, the sensitivity of the qPCR assay was ten times higher than LAMP. Our findings indicate that this LAMP method has a good sensitivity and high specificity for the detection of A. marginale and may have a potential application in the detection and differentiation of bovine anaplasmosis.
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spelling Development of a loop-mediated isothermal amplification (LAMP) assay for the detection of Anaplasma marginaleLAMPqPCRAnaplasma marginaleDetectionSensibilitySpecificityParasitemia generated by Anaplasma marginale causes significant losses in the cattle industry. A major constraint to the effective control and management of anaplasmosis in livestock is the lack of a rapid and reliable diagnostic test to identify the parasite and allow for immediate therapy. In the present study, we developed a novel DNA-based assay for the detection of A. marginale in bovine blood samples, using loop-mediated isothermal amplification (LAMP). DNA from six cattle and hemoparasite samples (Babesia bovis, Babesia bigemina, Anaplasma centrale and A. marginale) were tested for specificity, sensitivity and cross-reactions. The developed LAMP procedures were also confirmed and compared with the qPCR method. The same gene sequence (major surface protein 1b, msp1b) of A. marginale was used to design a set of primers for the LAMP and qPCR assays. The results showed that LAMP is specific, as no positive signal was observed for the other tested hemoparasites. However, the sensitivity of the qPCR assay was ten times higher than LAMP. Our findings indicate that this LAMP method has a good sensitivity and high specificity for the detection of A. marginale and may have a potential application in the detection and differentiation of bovine anaplasmosis.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Brazilian Agricultural Research CorporationUniv Estadual Paulista, Fac Ciencias Agr & Vet, Via Acesso Prof Paulo Donato Castellane, BR-14884900 Jaboticabal, SP, BrazilEmbrapa Pecuaria Sudeste, Rodovia Washington Luiz,Km 234,CP 339, BR-13560970 Sao Carlos, SP, BrazilCtr Pesquisa Genet & Reprod Anim, Inst Zootecnia, Rua Heitor Penteado 56, BR-13380011 Nova Odessa, SP, BrazilUniv Estadual Paulista, Fac Ciencias Agr & Vet, Via Acesso Prof Paulo Donato Castellane, BR-14884900 Jaboticabal, SP, BrazilFAPESP: 2016/07216-7FAPESP: 2017/11297-5CNPq: 153231/2018-1Brazilian Agricultural Research Corporation: Embrapa SEG 02.12.02.008.00.00SpringerUniversidade Estadual Paulista (Unesp)Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)Ctr Pesquisa Genet & Reprod AnimGiglioti, Rodrigo [UNESP]Bassetto, Cesar Cristiano [UNESP]Okino, Cintia HiromiOliveira, Henrique Nunes de [UNESP]Sena Oliveira, Marcia Cristina de2019-10-05T12:41:05Z2019-10-05T12:41:05Z2019-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article65-72http://dx.doi.org/10.1007/s10493-018-0327-yExperimental And Applied Acarology. Dordrecht: Springer, v. 77, n. 1, p. 65-72, 2019.0168-8162http://hdl.handle.net/11449/18663110.1007/s10493-018-0327-yWOS:000457939000006Web of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengExperimental And Applied Acarologyinfo:eu-repo/semantics/openAccess2024-06-07T18:40:13Zoai:repositorio.unesp.br:11449/186631Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T14:51:58.569413Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Development of a loop-mediated isothermal amplification (LAMP) assay for the detection of Anaplasma marginale
title Development of a loop-mediated isothermal amplification (LAMP) assay for the detection of Anaplasma marginale
spellingShingle Development of a loop-mediated isothermal amplification (LAMP) assay for the detection of Anaplasma marginale
Giglioti, Rodrigo [UNESP]
LAMP
qPCR
Anaplasma marginale
Detection
Sensibility
Specificity
title_short Development of a loop-mediated isothermal amplification (LAMP) assay for the detection of Anaplasma marginale
title_full Development of a loop-mediated isothermal amplification (LAMP) assay for the detection of Anaplasma marginale
title_fullStr Development of a loop-mediated isothermal amplification (LAMP) assay for the detection of Anaplasma marginale
title_full_unstemmed Development of a loop-mediated isothermal amplification (LAMP) assay for the detection of Anaplasma marginale
title_sort Development of a loop-mediated isothermal amplification (LAMP) assay for the detection of Anaplasma marginale
author Giglioti, Rodrigo [UNESP]
author_facet Giglioti, Rodrigo [UNESP]
Bassetto, Cesar Cristiano [UNESP]
Okino, Cintia Hiromi
Oliveira, Henrique Nunes de [UNESP]
Sena Oliveira, Marcia Cristina de
author_role author
author2 Bassetto, Cesar Cristiano [UNESP]
Okino, Cintia Hiromi
Oliveira, Henrique Nunes de [UNESP]
Sena Oliveira, Marcia Cristina de
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Universidade Estadual Paulista (Unesp)
Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)
Ctr Pesquisa Genet & Reprod Anim
dc.contributor.author.fl_str_mv Giglioti, Rodrigo [UNESP]
Bassetto, Cesar Cristiano [UNESP]
Okino, Cintia Hiromi
Oliveira, Henrique Nunes de [UNESP]
Sena Oliveira, Marcia Cristina de
dc.subject.por.fl_str_mv LAMP
qPCR
Anaplasma marginale
Detection
Sensibility
Specificity
topic LAMP
qPCR
Anaplasma marginale
Detection
Sensibility
Specificity
description Parasitemia generated by Anaplasma marginale causes significant losses in the cattle industry. A major constraint to the effective control and management of anaplasmosis in livestock is the lack of a rapid and reliable diagnostic test to identify the parasite and allow for immediate therapy. In the present study, we developed a novel DNA-based assay for the detection of A. marginale in bovine blood samples, using loop-mediated isothermal amplification (LAMP). DNA from six cattle and hemoparasite samples (Babesia bovis, Babesia bigemina, Anaplasma centrale and A. marginale) were tested for specificity, sensitivity and cross-reactions. The developed LAMP procedures were also confirmed and compared with the qPCR method. The same gene sequence (major surface protein 1b, msp1b) of A. marginale was used to design a set of primers for the LAMP and qPCR assays. The results showed that LAMP is specific, as no positive signal was observed for the other tested hemoparasites. However, the sensitivity of the qPCR assay was ten times higher than LAMP. Our findings indicate that this LAMP method has a good sensitivity and high specificity for the detection of A. marginale and may have a potential application in the detection and differentiation of bovine anaplasmosis.
publishDate 2019
dc.date.none.fl_str_mv 2019-10-05T12:41:05Z
2019-10-05T12:41:05Z
2019-01-01
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.uri.fl_str_mv http://dx.doi.org/10.1007/s10493-018-0327-y
Experimental And Applied Acarology. Dordrecht: Springer, v. 77, n. 1, p. 65-72, 2019.
0168-8162
http://hdl.handle.net/11449/186631
10.1007/s10493-018-0327-y
WOS:000457939000006
url http://dx.doi.org/10.1007/s10493-018-0327-y
http://hdl.handle.net/11449/186631
identifier_str_mv Experimental And Applied Acarology. Dordrecht: Springer, v. 77, n. 1, p. 65-72, 2019.
0168-8162
10.1007/s10493-018-0327-y
WOS:000457939000006
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Experimental And Applied Acarology
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 65-72
dc.publisher.none.fl_str_mv Springer
publisher.none.fl_str_mv Springer
dc.source.none.fl_str_mv Web of Science
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
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