ASSESSMENT OF PRIMERS DESIGNED FROM THE SMALL RIBOSOMAL SUBUNIT RNA FOR SPECIFIC DISCRIMINATION BETWEEN BABESIA BIGEMINA AND BABESIA BOVIS BY PCR
Autor(a) principal: | |
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Data de Publicação: | 2006 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | por |
Título da fonte: | Ciência animal brasileira (Online) |
Texto Completo: | https://revistas.ufg.br/vet/article/view/288 |
Resumo: | Six pairs of species-specific primers were designed from the alignment of the sequences of the SS rRNA gene obtained from the Genbank database for Babesia bigemina (accession number X59604) and for B. bovis (accession number U06105). Three pairs of primers were designed specifically for B. bigemina and another 3 sets of primers for B. bovis. All oligonucleotide sequences selected as primers were examined for similarities to other organisms through the Genbank Blast procedure and these 6 sets of primers demonstrated high level of specificity. The synthetic oligonucleotides were also tested for specificity by PCR assay using genomic DNA extracted from 40 isolates of B. bigemina and 30 from B. bovis, obtained in six different States of Brazil. All 6 sets of primers were validated as 100% specific for the respective parasite. The PCR amplified the expected fragments for each set of primers, as follows: a) B. bigemina: primers GAU5 forward/GAU6 reverse with amplicon of 1,124 bp; primers GAU5 forward/GAU8 reverse with amplicon of 458 bp; primers GAU7 forward/GAU6 reverse with amplicon of 685 bp; b) B. bovis: primers GAU9 forward/GAU10 reverse with amplicon of 541 bp; primers GAU9 forward/GAU 13 reverse with amplicon of 883 bp; primers SOGIN forward/GAU 10 with amplicon of 1211 bp. KEYWORDS: Babesia bovis, Babesia bigemina, SS rRNA gene, PCR, primers, bovine babesiosis. |
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Ciência animal brasileira (Online) |
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ASSESSMENT OF PRIMERS DESIGNED FROM THE SMALL RIBOSOMAL SUBUNIT RNA FOR SPECIFIC DISCRIMINATION BETWEEN BABESIA BIGEMINA AND BABESIA BOVIS BY PCRSix pairs of species-specific primers were designed from the alignment of the sequences of the SS rRNA gene obtained from the Genbank database for Babesia bigemina (accession number X59604) and for B. bovis (accession number U06105). Three pairs of primers were designed specifically for B. bigemina and another 3 sets of primers for B. bovis. All oligonucleotide sequences selected as primers were examined for similarities to other organisms through the Genbank Blast procedure and these 6 sets of primers demonstrated high level of specificity. The synthetic oligonucleotides were also tested for specificity by PCR assay using genomic DNA extracted from 40 isolates of B. bigemina and 30 from B. bovis, obtained in six different States of Brazil. All 6 sets of primers were validated as 100% specific for the respective parasite. The PCR amplified the expected fragments for each set of primers, as follows: a) B. bigemina: primers GAU5 forward/GAU6 reverse with amplicon of 1,124 bp; primers GAU5 forward/GAU8 reverse with amplicon of 458 bp; primers GAU7 forward/GAU6 reverse with amplicon of 685 bp; b) B. bovis: primers GAU9 forward/GAU10 reverse with amplicon of 541 bp; primers GAU9 forward/GAU 13 reverse with amplicon of 883 bp; primers SOGIN forward/GAU 10 with amplicon of 1211 bp. KEYWORDS: Babesia bovis, Babesia bigemina, SS rRNA gene, PCR, primers, bovine babesiosis.Universidade Federal de Goiás2006-10-25info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionAvaliado por paresapplication/pdfhttps://revistas.ufg.br/vet/article/view/28810.5216/cab.v3i2.288Brazilian Animal Science/ Ciência Animal Brasileira; Vol. 3 No. 2 (2002); 27-32Ciência Animal Brasileira / Brazilian Animal Science; v. 3 n. 2 (2002); 27-321809-68911518-2797reponame:Ciência animal brasileira (Online)instname:Universidade Federal de Goiás (UFG)instacron:UFGporhttps://revistas.ufg.br/vet/article/view/288/256Linhares, Guido Fontgalland CoelhoSantana, Angela PatríciaLaueman, Lloyd H.Madruga, Cláudio R.info:eu-repo/semantics/openAccess2013-10-12T20:38:51Zoai:ojs.revistas.ufg.br:article/288Revistahttps://revistas.ufg.br/vetPUBhttps://revistas.ufg.br/vet/oai||revistacab@gmail.com1809-68911518-2797opendoar:2013-10-12T20:38:51Ciência animal brasileira (Online) - Universidade Federal de Goiás (UFG)false |
dc.title.none.fl_str_mv |
ASSESSMENT OF PRIMERS DESIGNED FROM THE SMALL RIBOSOMAL SUBUNIT RNA FOR SPECIFIC DISCRIMINATION BETWEEN BABESIA BIGEMINA AND BABESIA BOVIS BY PCR |
title |
ASSESSMENT OF PRIMERS DESIGNED FROM THE SMALL RIBOSOMAL SUBUNIT RNA FOR SPECIFIC DISCRIMINATION BETWEEN BABESIA BIGEMINA AND BABESIA BOVIS BY PCR |
spellingShingle |
ASSESSMENT OF PRIMERS DESIGNED FROM THE SMALL RIBOSOMAL SUBUNIT RNA FOR SPECIFIC DISCRIMINATION BETWEEN BABESIA BIGEMINA AND BABESIA BOVIS BY PCR Linhares, Guido Fontgalland Coelho |
title_short |
ASSESSMENT OF PRIMERS DESIGNED FROM THE SMALL RIBOSOMAL SUBUNIT RNA FOR SPECIFIC DISCRIMINATION BETWEEN BABESIA BIGEMINA AND BABESIA BOVIS BY PCR |
title_full |
ASSESSMENT OF PRIMERS DESIGNED FROM THE SMALL RIBOSOMAL SUBUNIT RNA FOR SPECIFIC DISCRIMINATION BETWEEN BABESIA BIGEMINA AND BABESIA BOVIS BY PCR |
title_fullStr |
ASSESSMENT OF PRIMERS DESIGNED FROM THE SMALL RIBOSOMAL SUBUNIT RNA FOR SPECIFIC DISCRIMINATION BETWEEN BABESIA BIGEMINA AND BABESIA BOVIS BY PCR |
title_full_unstemmed |
ASSESSMENT OF PRIMERS DESIGNED FROM THE SMALL RIBOSOMAL SUBUNIT RNA FOR SPECIFIC DISCRIMINATION BETWEEN BABESIA BIGEMINA AND BABESIA BOVIS BY PCR |
title_sort |
ASSESSMENT OF PRIMERS DESIGNED FROM THE SMALL RIBOSOMAL SUBUNIT RNA FOR SPECIFIC DISCRIMINATION BETWEEN BABESIA BIGEMINA AND BABESIA BOVIS BY PCR |
author |
Linhares, Guido Fontgalland Coelho |
author_facet |
Linhares, Guido Fontgalland Coelho Santana, Angela Patrícia Laueman, Lloyd H. Madruga, Cláudio R. |
author_role |
author |
author2 |
Santana, Angela Patrícia Laueman, Lloyd H. Madruga, Cláudio R. |
author2_role |
author author author |
dc.contributor.author.fl_str_mv |
Linhares, Guido Fontgalland Coelho Santana, Angela Patrícia Laueman, Lloyd H. Madruga, Cláudio R. |
description |
Six pairs of species-specific primers were designed from the alignment of the sequences of the SS rRNA gene obtained from the Genbank database for Babesia bigemina (accession number X59604) and for B. bovis (accession number U06105). Three pairs of primers were designed specifically for B. bigemina and another 3 sets of primers for B. bovis. All oligonucleotide sequences selected as primers were examined for similarities to other organisms through the Genbank Blast procedure and these 6 sets of primers demonstrated high level of specificity. The synthetic oligonucleotides were also tested for specificity by PCR assay using genomic DNA extracted from 40 isolates of B. bigemina and 30 from B. bovis, obtained in six different States of Brazil. All 6 sets of primers were validated as 100% specific for the respective parasite. The PCR amplified the expected fragments for each set of primers, as follows: a) B. bigemina: primers GAU5 forward/GAU6 reverse with amplicon of 1,124 bp; primers GAU5 forward/GAU8 reverse with amplicon of 458 bp; primers GAU7 forward/GAU6 reverse with amplicon of 685 bp; b) B. bovis: primers GAU9 forward/GAU10 reverse with amplicon of 541 bp; primers GAU9 forward/GAU 13 reverse with amplicon of 883 bp; primers SOGIN forward/GAU 10 with amplicon of 1211 bp. KEYWORDS: Babesia bovis, Babesia bigemina, SS rRNA gene, PCR, primers, bovine babesiosis. |
publishDate |
2006 |
dc.date.none.fl_str_mv |
2006-10-25 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Avaliado por pares |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://revistas.ufg.br/vet/article/view/288 10.5216/cab.v3i2.288 |
url |
https://revistas.ufg.br/vet/article/view/288 |
identifier_str_mv |
10.5216/cab.v3i2.288 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.none.fl_str_mv |
https://revistas.ufg.br/vet/article/view/288/256 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Goiás |
publisher.none.fl_str_mv |
Universidade Federal de Goiás |
dc.source.none.fl_str_mv |
Brazilian Animal Science/ Ciência Animal Brasileira; Vol. 3 No. 2 (2002); 27-32 Ciência Animal Brasileira / Brazilian Animal Science; v. 3 n. 2 (2002); 27-32 1809-6891 1518-2797 reponame:Ciência animal brasileira (Online) instname:Universidade Federal de Goiás (UFG) instacron:UFG |
instname_str |
Universidade Federal de Goiás (UFG) |
instacron_str |
UFG |
institution |
UFG |
reponame_str |
Ciência animal brasileira (Online) |
collection |
Ciência animal brasileira (Online) |
repository.name.fl_str_mv |
Ciência animal brasileira (Online) - Universidade Federal de Goiás (UFG) |
repository.mail.fl_str_mv |
||revistacab@gmail.com |
_version_ |
1789440215441473536 |