Comparison of ovine β-globin haplotype sequences and a new multiplex PCR for identification
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.vetpar.2021.109592 http://hdl.handle.net/11449/223212 |
Resumo: | Deletion of pre-adult βC-globin in sheep harboring BB haplotype of β-globin was associated to decreased tolerance to anemia and hypoxia, and consequently, reduced resistance to Haemonchus contortus infection, which is closely related to severe anemia. Recently, a qPCR using hydrolysis probe was successfully developed for β-globin haplotype identification, and association between resistance against H. contortus and presence of βA allele was observed in Morada Nova sheep. Thus, this study aimed to better investigate the differences between β-globin haplotypes and to develop a conventional multiplex PCR, as an alternative to qPCR assay for β-globin haplotype identification. A total of 333 Morada Nova lambs had their blood collected and tested by both qPCR and new multiplex PCR, and 100 % of agreement was observed between the results. Since different primers were designed for such assay development, including different target genes, high specificity of both methods may be also highlighted. Three A haplotype samples were submitted to DNA Sanger sequencing of β-globin gene and compared to sequences previously deposited in Genbank. One nucleotide deletion in intronic region was observed only in AA haplotype of Morada Nova animals, while in BB animals the nucleotide remained present. To the best of our knowledge, this is the first report of multiplex conventional PCR for ovine β-globin haplotype identification. The advantages of the developed conventional PCR are reduced reagents costs (less than a half price) and wider reachability, since even labs without real time PCR thermocyclers are able to offer this assay. Therefore, it may become an important tool for sheep producers to improve genetic selection of parasite resistant animals. |
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Repositório Institucional da UNESP |
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Comparison of ovine β-globin haplotype sequences and a new multiplex PCR for identificationAlleleB haplotypeBeta globin CMorada NovaResistanceSheepDeletion of pre-adult βC-globin in sheep harboring BB haplotype of β-globin was associated to decreased tolerance to anemia and hypoxia, and consequently, reduced resistance to Haemonchus contortus infection, which is closely related to severe anemia. Recently, a qPCR using hydrolysis probe was successfully developed for β-globin haplotype identification, and association between resistance against H. contortus and presence of βA allele was observed in Morada Nova sheep. Thus, this study aimed to better investigate the differences between β-globin haplotypes and to develop a conventional multiplex PCR, as an alternative to qPCR assay for β-globin haplotype identification. A total of 333 Morada Nova lambs had their blood collected and tested by both qPCR and new multiplex PCR, and 100 % of agreement was observed between the results. Since different primers were designed for such assay development, including different target genes, high specificity of both methods may be also highlighted. Three A haplotype samples were submitted to DNA Sanger sequencing of β-globin gene and compared to sequences previously deposited in Genbank. One nucleotide deletion in intronic region was observed only in AA haplotype of Morada Nova animals, while in BB animals the nucleotide remained present. To the best of our knowledge, this is the first report of multiplex conventional PCR for ovine β-globin haplotype identification. The advantages of the developed conventional PCR are reduced reagents costs (less than a half price) and wider reachability, since even labs without real time PCR thermocyclers are able to offer this assay. Therefore, it may become an important tool for sheep producers to improve genetic selection of parasite resistant animals.Embrapa Pecuária Sudeste, Rodovia Washington Luiz ,Km 234 s/n ,PO Box 339Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista (UNESP), Via de Acesso Prof. Paulo Donato Castellane ,s/nFaculdade de Ciências Agrárias e Veterinárias Universidade Estadual Paulista (UNESP), Via de Acesso Prof. Paulo Donato Castellane ,s/nEmpresa Brasileira de Pesquisa Agropecuária (EMBRAPA)Universidade Estadual Paulista (UNESP)Okino, Cintia HiromiCaminotto, Lara Cristina FrizoniKapritchkoff, Rafaela Tami Ikeda [UNESP]Niciura, Simone Cristina MéoEsteves, Sérgio NovitaSantos, Isabella Barbosa Dos [UNESP]Toscano, João Henrique Barbosa [UNESP]Oliveira, M. C.S.Chagas, A. C.S.2022-04-28T19:49:24Z2022-04-28T19:49:24Z2021-12-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article109592http://dx.doi.org/10.1016/j.vetpar.2021.109592Veterinary parasitology, v. 300, p. 109592-.1873-2550http://hdl.handle.net/11449/22321210.1016/j.vetpar.2021.1095922-s2.0-85122546745Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengVeterinary parasitologyinfo:eu-repo/semantics/openAccess2022-04-28T19:49:24Zoai:repositorio.unesp.br:11449/223212Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462022-04-28T19:49:24Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Comparison of ovine β-globin haplotype sequences and a new multiplex PCR for identification |
title |
Comparison of ovine β-globin haplotype sequences and a new multiplex PCR for identification |
spellingShingle |
Comparison of ovine β-globin haplotype sequences and a new multiplex PCR for identification Okino, Cintia Hiromi Allele B haplotype Beta globin C Morada Nova Resistance Sheep |
title_short |
Comparison of ovine β-globin haplotype sequences and a new multiplex PCR for identification |
title_full |
Comparison of ovine β-globin haplotype sequences and a new multiplex PCR for identification |
title_fullStr |
Comparison of ovine β-globin haplotype sequences and a new multiplex PCR for identification |
title_full_unstemmed |
Comparison of ovine β-globin haplotype sequences and a new multiplex PCR for identification |
title_sort |
Comparison of ovine β-globin haplotype sequences and a new multiplex PCR for identification |
author |
Okino, Cintia Hiromi |
author_facet |
Okino, Cintia Hiromi Caminotto, Lara Cristina Frizoni Kapritchkoff, Rafaela Tami Ikeda [UNESP] Niciura, Simone Cristina Méo Esteves, Sérgio Novita Santos, Isabella Barbosa Dos [UNESP] Toscano, João Henrique Barbosa [UNESP] Oliveira, M. C.S. Chagas, A. C.S. |
author_role |
author |
author2 |
Caminotto, Lara Cristina Frizoni Kapritchkoff, Rafaela Tami Ikeda [UNESP] Niciura, Simone Cristina Méo Esteves, Sérgio Novita Santos, Isabella Barbosa Dos [UNESP] Toscano, João Henrique Barbosa [UNESP] Oliveira, M. C.S. Chagas, A. C.S. |
author2_role |
author author author author author author author author |
dc.contributor.none.fl_str_mv |
Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) Universidade Estadual Paulista (UNESP) |
dc.contributor.author.fl_str_mv |
Okino, Cintia Hiromi Caminotto, Lara Cristina Frizoni Kapritchkoff, Rafaela Tami Ikeda [UNESP] Niciura, Simone Cristina Méo Esteves, Sérgio Novita Santos, Isabella Barbosa Dos [UNESP] Toscano, João Henrique Barbosa [UNESP] Oliveira, M. C.S. Chagas, A. C.S. |
dc.subject.por.fl_str_mv |
Allele B haplotype Beta globin C Morada Nova Resistance Sheep |
topic |
Allele B haplotype Beta globin C Morada Nova Resistance Sheep |
description |
Deletion of pre-adult βC-globin in sheep harboring BB haplotype of β-globin was associated to decreased tolerance to anemia and hypoxia, and consequently, reduced resistance to Haemonchus contortus infection, which is closely related to severe anemia. Recently, a qPCR using hydrolysis probe was successfully developed for β-globin haplotype identification, and association between resistance against H. contortus and presence of βA allele was observed in Morada Nova sheep. Thus, this study aimed to better investigate the differences between β-globin haplotypes and to develop a conventional multiplex PCR, as an alternative to qPCR assay for β-globin haplotype identification. A total of 333 Morada Nova lambs had their blood collected and tested by both qPCR and new multiplex PCR, and 100 % of agreement was observed between the results. Since different primers were designed for such assay development, including different target genes, high specificity of both methods may be also highlighted. Three A haplotype samples were submitted to DNA Sanger sequencing of β-globin gene and compared to sequences previously deposited in Genbank. One nucleotide deletion in intronic region was observed only in AA haplotype of Morada Nova animals, while in BB animals the nucleotide remained present. To the best of our knowledge, this is the first report of multiplex conventional PCR for ovine β-globin haplotype identification. The advantages of the developed conventional PCR are reduced reagents costs (less than a half price) and wider reachability, since even labs without real time PCR thermocyclers are able to offer this assay. Therefore, it may become an important tool for sheep producers to improve genetic selection of parasite resistant animals. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-12-01 2022-04-28T19:49:24Z 2022-04-28T19:49:24Z |
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.vetpar.2021.109592 Veterinary parasitology, v. 300, p. 109592-. 1873-2550 http://hdl.handle.net/11449/223212 10.1016/j.vetpar.2021.109592 2-s2.0-85122546745 |
url |
http://dx.doi.org/10.1016/j.vetpar.2021.109592 http://hdl.handle.net/11449/223212 |
identifier_str_mv |
Veterinary parasitology, v. 300, p. 109592-. 1873-2550 10.1016/j.vetpar.2021.109592 2-s2.0-85122546745 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Veterinary parasitology |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
109592 |
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_ |
1799964842825613312 |