DNA extraction methods for molecular detection of Eimeria spp. in cattle and sheep
Autor(a) principal: | |
---|---|
Data de Publicação: | 2020 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Pesquisa Veterinária Brasileira (Online) |
Texto Completo: | http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-736X2020000700514 |
Resumo: | ABSTRACT: Molecular detection of Eimeria species in fecal samples can be useful for experimental and diagnostic purposes. However, the parasite quantity presence in feces and the oocyst wall are an obstacle in DNA extraction protocols. Therefore, adequate sampling and effective disruption of the oocysts are essential to improve the accuracy of DNA detection by PCR. The aims of this study were to evaluate the suitability of six protocols for DNA extraction from Eimeria spp. present in bovine and sheep. Twenty pools of fecal samples from cattle (10 pools) and sheep (10 pools) were distributed to six DNA extraction protocols: commercial kit, commercial kit with modification, DNAzol, cetyl-trimethyl ammonium bromide (CTAB), glass beads and commercial kit for fecal samples. Fecal samples were submitted to DNA extraction and PCR. Among the protocols tested, CTAB was determined to be most suitable for DNA extraction from oocysts (90% of DNA detection by PCR); DNAzol and CTAB resulted in higher DNA detection from bovine samples (80%). CTAB and commercial kit with modification improved PCR detection of Eimeria spp. in sheep samples, with positive amplification of DNA in all tested samples. |
id |
EMBRAPA-2_b5cc38d5f1c789fb6d10650b003ca107 |
---|---|
oai_identifier_str |
oai:scielo:S0100-736X2020000700514 |
network_acronym_str |
EMBRAPA-2 |
network_name_str |
Pesquisa Veterinária Brasileira (Online) |
repository_id_str |
|
spelling |
DNA extraction methods for molecular detection of Eimeria spp. in cattle and sheepDNA extractionmolecular detectionEimeria spp.cattlesheepPCRfecal samplesABSTRACT: Molecular detection of Eimeria species in fecal samples can be useful for experimental and diagnostic purposes. However, the parasite quantity presence in feces and the oocyst wall are an obstacle in DNA extraction protocols. Therefore, adequate sampling and effective disruption of the oocysts are essential to improve the accuracy of DNA detection by PCR. The aims of this study were to evaluate the suitability of six protocols for DNA extraction from Eimeria spp. present in bovine and sheep. Twenty pools of fecal samples from cattle (10 pools) and sheep (10 pools) were distributed to six DNA extraction protocols: commercial kit, commercial kit with modification, DNAzol, cetyl-trimethyl ammonium bromide (CTAB), glass beads and commercial kit for fecal samples. Fecal samples were submitted to DNA extraction and PCR. Among the protocols tested, CTAB was determined to be most suitable for DNA extraction from oocysts (90% of DNA detection by PCR); DNAzol and CTAB resulted in higher DNA detection from bovine samples (80%). CTAB and commercial kit with modification improved PCR detection of Eimeria spp. in sheep samples, with positive amplification of DNA in all tested samples.Colégio Brasileiro de Patologia Animal - CBPA2020-07-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-736X2020000700514Pesquisa Veterinária Brasileira v.40 n.7 2020reponame:Pesquisa Veterinária Brasileira (Online)instname:Colégio Brasileiro de Patologia Animal (CBPA)instacron:EMBRAPA10.1590/1678-5150-pvb-6625info:eu-repo/semantics/openAccessReginato,Caroline Z.Bräunig,PatríciaPortella,Luiza P.Mortari,Ana Paula G.Minuzzi,Camila E.Sangioni,Luis AntonioVogel,Fernanda S.F.eng2020-09-16T00:00:00Zoai:scielo:S0100-736X2020000700514Revistahttp://www.pvb.com.br/https://old.scielo.br/oai/scielo-oai.phpcolegio@cbpa.org.br||pvb@pvb.com.br0100-736X1678-5150opendoar:2020-09-16T00:00Pesquisa Veterinária Brasileira (Online) - Colégio Brasileiro de Patologia Animal (CBPA)false |
dc.title.none.fl_str_mv |
DNA extraction methods for molecular detection of Eimeria spp. in cattle and sheep |
title |
DNA extraction methods for molecular detection of Eimeria spp. in cattle and sheep |
spellingShingle |
DNA extraction methods for molecular detection of Eimeria spp. in cattle and sheep Reginato,Caroline Z. DNA extraction molecular detection Eimeria spp. cattle sheep PCR fecal samples |
title_short |
DNA extraction methods for molecular detection of Eimeria spp. in cattle and sheep |
title_full |
DNA extraction methods for molecular detection of Eimeria spp. in cattle and sheep |
title_fullStr |
DNA extraction methods for molecular detection of Eimeria spp. in cattle and sheep |
title_full_unstemmed |
DNA extraction methods for molecular detection of Eimeria spp. in cattle and sheep |
title_sort |
DNA extraction methods for molecular detection of Eimeria spp. in cattle and sheep |
author |
Reginato,Caroline Z. |
author_facet |
Reginato,Caroline Z. Bräunig,Patrícia Portella,Luiza P. Mortari,Ana Paula G. Minuzzi,Camila E. Sangioni,Luis Antonio Vogel,Fernanda S.F. |
author_role |
author |
author2 |
Bräunig,Patrícia Portella,Luiza P. Mortari,Ana Paula G. Minuzzi,Camila E. Sangioni,Luis Antonio Vogel,Fernanda S.F. |
author2_role |
author author author author author author |
dc.contributor.author.fl_str_mv |
Reginato,Caroline Z. Bräunig,Patrícia Portella,Luiza P. Mortari,Ana Paula G. Minuzzi,Camila E. Sangioni,Luis Antonio Vogel,Fernanda S.F. |
dc.subject.por.fl_str_mv |
DNA extraction molecular detection Eimeria spp. cattle sheep PCR fecal samples |
topic |
DNA extraction molecular detection Eimeria spp. cattle sheep PCR fecal samples |
description |
ABSTRACT: Molecular detection of Eimeria species in fecal samples can be useful for experimental and diagnostic purposes. However, the parasite quantity presence in feces and the oocyst wall are an obstacle in DNA extraction protocols. Therefore, adequate sampling and effective disruption of the oocysts are essential to improve the accuracy of DNA detection by PCR. The aims of this study were to evaluate the suitability of six protocols for DNA extraction from Eimeria spp. present in bovine and sheep. Twenty pools of fecal samples from cattle (10 pools) and sheep (10 pools) were distributed to six DNA extraction protocols: commercial kit, commercial kit with modification, DNAzol, cetyl-trimethyl ammonium bromide (CTAB), glass beads and commercial kit for fecal samples. Fecal samples were submitted to DNA extraction and PCR. Among the protocols tested, CTAB was determined to be most suitable for DNA extraction from oocysts (90% of DNA detection by PCR); DNAzol and CTAB resulted in higher DNA detection from bovine samples (80%). CTAB and commercial kit with modification improved PCR detection of Eimeria spp. in sheep samples, with positive amplification of DNA in all tested samples. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-07-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-736X2020000700514 |
url |
http://old.scielo.br/scielo.php?script=sci_arttext&pid=S0100-736X2020000700514 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/1678-5150-pvb-6625 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Colégio Brasileiro de Patologia Animal - CBPA |
publisher.none.fl_str_mv |
Colégio Brasileiro de Patologia Animal - CBPA |
dc.source.none.fl_str_mv |
Pesquisa Veterinária Brasileira v.40 n.7 2020 reponame:Pesquisa Veterinária Brasileira (Online) instname:Colégio Brasileiro de Patologia Animal (CBPA) instacron:EMBRAPA |
instname_str |
Colégio Brasileiro de Patologia Animal (CBPA) |
instacron_str |
EMBRAPA |
institution |
EMBRAPA |
reponame_str |
Pesquisa Veterinária Brasileira (Online) |
collection |
Pesquisa Veterinária Brasileira (Online) |
repository.name.fl_str_mv |
Pesquisa Veterinária Brasileira (Online) - Colégio Brasileiro de Patologia Animal (CBPA) |
repository.mail.fl_str_mv |
colegio@cbpa.org.br||pvb@pvb.com.br |
_version_ |
1754122240402653184 |