Semi-quantitative evaluation of Babesia bovis and B. bigemina infection levels estimated by HRM analysis
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Outros Autores: | , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1016/j.ttbdis.2021.101753 http://hdl.handle.net/11449/206466 |
Resumo: | Bovine babesiosis is economically the most important arthropod-borne disease of cattle worldwide. The most significant damage caused by bovine babesiosis is attributed to Babesia bovis due to its higher pathogenicity. This study aimed to develop a real-time PCR method followed by HRM (high-resolution melting) analysis for the simultaneous detection of B. bovis and B. bigemina, enabling a semi-quantitative analysis of Babesia levels using a single-tube reaction. The HRM was compared with real-time PCR using species-specific hydrolysis probes. The HRM analysis allowed to differentiate both Babesia species and was sensitive in the detection and differentiation of 10% for each Babesia species in the sample. Our results suggest the use of this method to estimate the prevalence of infections by B. bovis or B. bigemina as an alternative to the methods of absolute quantification by real-time PCR since it neither requires precise estimates of the number of DNA loads nor the construction of calibration curves. The simultaneous detection of the two Babesia species can be used to characterise the infection levels in cattle populations from different geographical regions, allowing a better control of these diseases. |
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Semi-quantitative evaluation of Babesia bovis and B. bigemina infection levels estimated by HRM analysisBabesiosisHRM analysisqPCRsemi-quantificationBovine babesiosis is economically the most important arthropod-borne disease of cattle worldwide. The most significant damage caused by bovine babesiosis is attributed to Babesia bovis due to its higher pathogenicity. This study aimed to develop a real-time PCR method followed by HRM (high-resolution melting) analysis for the simultaneous detection of B. bovis and B. bigemina, enabling a semi-quantitative analysis of Babesia levels using a single-tube reaction. The HRM was compared with real-time PCR using species-specific hydrolysis probes. The HRM analysis allowed to differentiate both Babesia species and was sensitive in the detection and differentiation of 10% for each Babesia species in the sample. Our results suggest the use of this method to estimate the prevalence of infections by B. bovis or B. bigemina as an alternative to the methods of absolute quantification by real-time PCR since it neither requires precise estimates of the number of DNA loads nor the construction of calibration curves. The simultaneous detection of the two Babesia species can be used to characterise the infection levels in cattle populations from different geographical regions, allowing a better control of these diseases.National Institute of Animal ScienceInstituto de Zootecnia Rua Heitor Penteado, n. 56, Nova OdessaEmbrapa Pecuária SudesteUniversidade Estadual Júlio de Mesquita FilhoUniversidade Estadual Júlio de Mesquita FilhoRua Heitor PenteadoEmpresa Brasileira de Pesquisa Agropecuária (EMBRAPA)Universidade Estadual Paulista (Unesp)Giglioti, RodrigoOkino, Cintia HiromiAzevedo, Bianca TaináWedy, Bruna Costa RodriguesGutmanis, GuntaVeríssimo, Cecília JoséKatiki, Luciana MoritaFilho, Anibal Eugênio Verceside Oliveira, Henrique Nunes [UNESP]Oliveira, Márcia Cristina de Sena2021-06-25T10:32:32Z2021-06-25T10:32:32Z2021-09-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlehttp://dx.doi.org/10.1016/j.ttbdis.2021.101753Ticks and Tick-borne Diseases, v. 12, n. 5, 2021.1877-96031877-959Xhttp://hdl.handle.net/11449/20646610.1016/j.ttbdis.2021.1017532-s2.0-85107718553Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengTicks and Tick-borne Diseasesinfo:eu-repo/semantics/openAccess2021-10-23T05:25:55Zoai:repositorio.unesp.br:11449/206466Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462021-10-23T05:25:55Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Semi-quantitative evaluation of Babesia bovis and B. bigemina infection levels estimated by HRM analysis |
title |
Semi-quantitative evaluation of Babesia bovis and B. bigemina infection levels estimated by HRM analysis |
spellingShingle |
Semi-quantitative evaluation of Babesia bovis and B. bigemina infection levels estimated by HRM analysis Giglioti, Rodrigo Babesiosis HRM analysis qPCR semi-quantification |
title_short |
Semi-quantitative evaluation of Babesia bovis and B. bigemina infection levels estimated by HRM analysis |
title_full |
Semi-quantitative evaluation of Babesia bovis and B. bigemina infection levels estimated by HRM analysis |
title_fullStr |
Semi-quantitative evaluation of Babesia bovis and B. bigemina infection levels estimated by HRM analysis |
title_full_unstemmed |
Semi-quantitative evaluation of Babesia bovis and B. bigemina infection levels estimated by HRM analysis |
title_sort |
Semi-quantitative evaluation of Babesia bovis and B. bigemina infection levels estimated by HRM analysis |
author |
Giglioti, Rodrigo |
author_facet |
Giglioti, Rodrigo Okino, Cintia Hiromi Azevedo, Bianca Tainá Wedy, Bruna Costa Rodrigues Gutmanis, Gunta Veríssimo, Cecília José Katiki, Luciana Morita Filho, Anibal Eugênio Vercesi de Oliveira, Henrique Nunes [UNESP] Oliveira, Márcia Cristina de Sena |
author_role |
author |
author2 |
Okino, Cintia Hiromi Azevedo, Bianca Tainá Wedy, Bruna Costa Rodrigues Gutmanis, Gunta Veríssimo, Cecília José Katiki, Luciana Morita Filho, Anibal Eugênio Vercesi de Oliveira, Henrique Nunes [UNESP] Oliveira, Márcia Cristina de Sena |
author2_role |
author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Rua Heitor Penteado Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
Giglioti, Rodrigo Okino, Cintia Hiromi Azevedo, Bianca Tainá Wedy, Bruna Costa Rodrigues Gutmanis, Gunta Veríssimo, Cecília José Katiki, Luciana Morita Filho, Anibal Eugênio Vercesi de Oliveira, Henrique Nunes [UNESP] Oliveira, Márcia Cristina de Sena |
dc.subject.por.fl_str_mv |
Babesiosis HRM analysis qPCR semi-quantification |
topic |
Babesiosis HRM analysis qPCR semi-quantification |
description |
Bovine babesiosis is economically the most important arthropod-borne disease of cattle worldwide. The most significant damage caused by bovine babesiosis is attributed to Babesia bovis due to its higher pathogenicity. This study aimed to develop a real-time PCR method followed by HRM (high-resolution melting) analysis for the simultaneous detection of B. bovis and B. bigemina, enabling a semi-quantitative analysis of Babesia levels using a single-tube reaction. The HRM was compared with real-time PCR using species-specific hydrolysis probes. The HRM analysis allowed to differentiate both Babesia species and was sensitive in the detection and differentiation of 10% for each Babesia species in the sample. Our results suggest the use of this method to estimate the prevalence of infections by B. bovis or B. bigemina as an alternative to the methods of absolute quantification by real-time PCR since it neither requires precise estimates of the number of DNA loads nor the construction of calibration curves. The simultaneous detection of the two Babesia species can be used to characterise the infection levels in cattle populations from different geographical regions, allowing a better control of these diseases. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-06-25T10:32:32Z 2021-06-25T10:32:32Z 2021-09-01 |
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/10.1016/j.ttbdis.2021.101753 Ticks and Tick-borne Diseases, v. 12, n. 5, 2021. 1877-9603 1877-959X http://hdl.handle.net/11449/206466 10.1016/j.ttbdis.2021.101753 2-s2.0-85107718553 |
url |
http://dx.doi.org/10.1016/j.ttbdis.2021.101753 http://hdl.handle.net/11449/206466 |
identifier_str_mv |
Ticks and Tick-borne Diseases, v. 12, n. 5, 2021. 1877-9603 1877-959X 10.1016/j.ttbdis.2021.101753 2-s2.0-85107718553 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Ticks and Tick-borne Diseases |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.source.none.fl_str_mv |
Scopus 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 |
|
_version_ |
1799965427790512128 |