Development of a polymerase chain reaction and its comparison with agar gel immunodiffusion test in the detection of bovine leukemia virus infection

Detalhes bibliográficos
Autor(a) principal: Camargos, Marcelo Fernandes
Data de Publicação: 2003
Outros Autores: Stancek, Daniel, Lessa, Leandro Moreira, Reis, Jenner Karlisson Pimenta, Rocha, Maurílio Andrade, Leite, Rômulo Cerqueira
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Brazilian Journal of Veterinary Research and Animal Science
Texto Completo: https://www.revistas.usp.br/bjvras/article/view/11311
Resumo: Polymerase chain reaction (PCR) was used for bovine leukemia virus (BLV) detection in the peripheral leukocytes of the infected bovines. The primers used were designed to amplify a part of env gene of BLV. PCR products were analyzed by agarose gel electrophoresis stained by ethidium bromide. The analytical specificity of PCR was confirmed by enzymatic restriction analysis of the PCR product with Bam HI and also by nucleotide sequence analysis of three PCR samples. Sixty five animals were tested for anti-BLV antibody, by agar gel-immunodiffusion test (AGID) and for direct BLV detection by PCR. There was a 73.80% concordance rate between the two tests. Four animals positive in AGID were PCR negative, while 13 AGID negative animals were found PCR positive. PCR got a 0.87 diagnosis sensitivity and 0.62 specificity. The developed PCR may be complementary tool in the diagnosis of BLV infection, but should have it diagnosis sensitivity improved.
id USP-49_4083fb14e6faff1b5006b1bbda577698
oai_identifier_str oai:revistas.usp.br:article/11311
network_acronym_str USP-49
network_name_str Brazilian Journal of Veterinary Research and Animal Science
repository_id_str https://www.revistas.usp.br/bjvras/index
spelling Development of a polymerase chain reaction and its comparison with agar gel immunodiffusion test in the detection of bovine leukemia virus infectionDesenvolvimento de uma reação em cadeia pela polimerase e comparação com a imunodifusão em gel de agar na detecção de infecções pelo vírus da leucemia bovinaImunodifusãoVírus da leucemia bovinaGenesReação em cadeia pela polimeraseImmunodiffusionBovine leukemia virusGenePolymerase chain reactionPolymerase chain reaction (PCR) was used for bovine leukemia virus (BLV) detection in the peripheral leukocytes of the infected bovines. The primers used were designed to amplify a part of env gene of BLV. PCR products were analyzed by agarose gel electrophoresis stained by ethidium bromide. The analytical specificity of PCR was confirmed by enzymatic restriction analysis of the PCR product with Bam HI and also by nucleotide sequence analysis of three PCR samples. Sixty five animals were tested for anti-BLV antibody, by agar gel-immunodiffusion test (AGID) and for direct BLV detection by PCR. There was a 73.80% concordance rate between the two tests. Four animals positive in AGID were PCR negative, while 13 AGID negative animals were found PCR positive. PCR got a 0.87 diagnosis sensitivity and 0.62 specificity. The developed PCR may be complementary tool in the diagnosis of BLV infection, but should have it diagnosis sensitivity improved.A reação em cadeia pela polimerase (PCR) foi utilizada para a detecção do vírus da leucemia bovina (VLB) em leucócitos periféricos de bovinos infectados. Os iniciadores utilizados foram construídos para amplificar uma parte do gene env do VLB. Os produtos da PCR foram analisados por eletroforese em gel de agarose corados por brometo de etídeo. A especificidade analítica da PCR foi confirmada por restrição enzimática dos produtos da reação com Bam HI e também pela análise da seqüência de três amostras. Sessenta e cinco animais foram testados para a presença de anticorpos anti-VLB, pela imunodifusão em gel de agar (IDGA) e pela PCR, para detecção direta do VLB. Houve 73,80% de concordância entre os dois testes. Quatro animais positivos na IDGA foram PCR negativos, enquanto 13 animais negativos na IDGA foram positivos na PCR. A sensibilidade diagnóstica obtida foi de 0,87 e a especificidade diagnóstica 0,62. A PCR desenvolvida pode ser uma ferramenta complementar no diagnóstico de infecções causadas pelo VLB, mas deve ter sua sensibilidade diagnóstica melhorada.Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia2003-01-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://www.revistas.usp.br/bjvras/article/view/1131110.1590/S1413-95962003000500005Brazilian Journal of Veterinary Research and Animal Science; Vol. 40 Núm. 5 (2003); 341-348Brazilian Journal of Veterinary Research and Animal Science; Vol. 40 No. 5 (2003); 341-348Brazilian Journal of Veterinary Research and Animal Science; v. 40 n. 5 (2003); 341-348Brazilian Journal of Veterinary Research and Animal Science; V. 40 N. 5 (2003); 341-3481678-44561413-9596reponame:Brazilian Journal of Veterinary Research and Animal Scienceinstname:Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP)instacron:USPenghttps://www.revistas.usp.br/bjvras/article/view/11311/13079Camargos, Marcelo FernandesStancek, DanielLessa, Leandro MoreiraReis, Jenner Karlisson PimentaRocha, Maurílio AndradeLeite, Rômulo Cerqueirainfo:eu-repo/semantics/openAccess2020-06-23T04:35:30Zoai:revistas.usp.br:article/11311Revistahttps://www.revistas.usp.br/bjvrasPUBhttps://www.revistas.usp.br/bjvras/oaibjvras@usp.br1413-95961413-9596opendoar:https://www.revistas.usp.br/bjvras/index2023-01-12T16:42:38.177967Brazilian Journal of Veterinary Research and Animal Science - Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP)false
dc.title.none.fl_str_mv Development of a polymerase chain reaction and its comparison with agar gel immunodiffusion test in the detection of bovine leukemia virus infection
Desenvolvimento de uma reação em cadeia pela polimerase e comparação com a imunodifusão em gel de agar na detecção de infecções pelo vírus da leucemia bovina
title Development of a polymerase chain reaction and its comparison with agar gel immunodiffusion test in the detection of bovine leukemia virus infection
spellingShingle Development of a polymerase chain reaction and its comparison with agar gel immunodiffusion test in the detection of bovine leukemia virus infection
Camargos, Marcelo Fernandes
Imunodifusão
Vírus da leucemia bovina
Genes
Reação em cadeia pela polimerase
Immunodiffusion
Bovine leukemia virus
Gene
Polymerase chain reaction
title_short Development of a polymerase chain reaction and its comparison with agar gel immunodiffusion test in the detection of bovine leukemia virus infection
title_full Development of a polymerase chain reaction and its comparison with agar gel immunodiffusion test in the detection of bovine leukemia virus infection
title_fullStr Development of a polymerase chain reaction and its comparison with agar gel immunodiffusion test in the detection of bovine leukemia virus infection
title_full_unstemmed Development of a polymerase chain reaction and its comparison with agar gel immunodiffusion test in the detection of bovine leukemia virus infection
title_sort Development of a polymerase chain reaction and its comparison with agar gel immunodiffusion test in the detection of bovine leukemia virus infection
author Camargos, Marcelo Fernandes
author_facet Camargos, Marcelo Fernandes
Stancek, Daniel
Lessa, Leandro Moreira
Reis, Jenner Karlisson Pimenta
Rocha, Maurílio Andrade
Leite, Rômulo Cerqueira
author_role author
author2 Stancek, Daniel
Lessa, Leandro Moreira
Reis, Jenner Karlisson Pimenta
Rocha, Maurílio Andrade
Leite, Rômulo Cerqueira
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Camargos, Marcelo Fernandes
Stancek, Daniel
Lessa, Leandro Moreira
Reis, Jenner Karlisson Pimenta
Rocha, Maurílio Andrade
Leite, Rômulo Cerqueira
dc.subject.por.fl_str_mv Imunodifusão
Vírus da leucemia bovina
Genes
Reação em cadeia pela polimerase
Immunodiffusion
Bovine leukemia virus
Gene
Polymerase chain reaction
topic Imunodifusão
Vírus da leucemia bovina
Genes
Reação em cadeia pela polimerase
Immunodiffusion
Bovine leukemia virus
Gene
Polymerase chain reaction
description Polymerase chain reaction (PCR) was used for bovine leukemia virus (BLV) detection in the peripheral leukocytes of the infected bovines. The primers used were designed to amplify a part of env gene of BLV. PCR products were analyzed by agarose gel electrophoresis stained by ethidium bromide. The analytical specificity of PCR was confirmed by enzymatic restriction analysis of the PCR product with Bam HI and also by nucleotide sequence analysis of three PCR samples. Sixty five animals were tested for anti-BLV antibody, by agar gel-immunodiffusion test (AGID) and for direct BLV detection by PCR. There was a 73.80% concordance rate between the two tests. Four animals positive in AGID were PCR negative, while 13 AGID negative animals were found PCR positive. PCR got a 0.87 diagnosis sensitivity and 0.62 specificity. The developed PCR may be complementary tool in the diagnosis of BLV infection, but should have it diagnosis sensitivity improved.
publishDate 2003
dc.date.none.fl_str_mv 2003-01-01
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://www.revistas.usp.br/bjvras/article/view/11311
10.1590/S1413-95962003000500005
url https://www.revistas.usp.br/bjvras/article/view/11311
identifier_str_mv 10.1590/S1413-95962003000500005
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv https://www.revistas.usp.br/bjvras/article/view/11311/13079
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 de São Paulo. Faculdade de Medicina Veterinária e Zootecnia
publisher.none.fl_str_mv Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia
dc.source.none.fl_str_mv Brazilian Journal of Veterinary Research and Animal Science; Vol. 40 Núm. 5 (2003); 341-348
Brazilian Journal of Veterinary Research and Animal Science; Vol. 40 No. 5 (2003); 341-348
Brazilian Journal of Veterinary Research and Animal Science; v. 40 n. 5 (2003); 341-348
Brazilian Journal of Veterinary Research and Animal Science; V. 40 N. 5 (2003); 341-348
1678-4456
1413-9596
reponame:Brazilian Journal of Veterinary Research and Animal Science
instname:Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP)
instacron:USP
instname_str Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP)
instacron_str USP
institution USP
reponame_str Brazilian Journal of Veterinary Research and Animal Science
collection Brazilian Journal of Veterinary Research and Animal Science
repository.name.fl_str_mv Brazilian Journal of Veterinary Research and Animal Science - Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo (FMVZ-USP)
repository.mail.fl_str_mv bjvras@usp.br
_version_ 1797051556719230976