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Registro Completo |
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Biblioteca(s): |
Embrapa Caprinos e Ovinos. |
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Data corrente: |
24/06/2013 |
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Data da última atualização: |
27/06/2016 |
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Autoria: |
OLIVEIRA, M. M. M.; MELO, M. A. de; ANDRADE, P. P. de; GOMES, S. M.; CAMPOS, A. C. |
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Título: |
Western blot para o diagnóstico das infecções pelos lentivírus de pequenos ruminantes em caprinos: um método simples para a produção de antígeno. |
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Ano de publicação: |
2013 |
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Fonte/Imprenta: |
Arquivo Brasileiro de Medicina Veterinária e Zootecnia, v. 65, n. 3, p. 694-698, 2013. |
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Idioma: |
Português |
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Conteúdo: |
Resumo: O sorodiagnóstico das lentiviroses de caprinos e ovinos é realizado principalmente pela imunodifusão em gel de agar (AGID) e/ou ELISA. Embora relativamente simples, esses métodos não identificam os antígenos virais reconhecidos na resposta imune do animal examinado, por isso o western blot (WB) vem ganhando maior relevância como ferramenta de diagnóstico dessas enfermidades. Neste trabalho, o antígeno utilizado no WB foi obtido através de um sistema simplificado de purificação: concentração por diálise do sobrenadante de culturas celulares infectadas, seguido de centrifugação em gradiente contínuo de sacarose. A separação das proteínas virais foi obtida por SDS-PAGE a 10% e a transferência para membranas de nitrocelulose realizada pelo sistema semi-úmido. A revelação das membranas mostrou reconhecimento pelo soro padrão positivo de cinco proteínas, com pesos moleculares de 14-16, 25, 40, 50 e 70 kDa. Todas as 8 amostras de soro caprino, positivas na ADIG, reconheceram pelo menos uma banda proteíca no immunoblot, variando contudo o número de bandas reconhecidas. Reação positiva à glícoproteína 40 (gp 40) foi observada em quatro animais, com intensidade de reação discreta em três deles. Dois animais apresentaram reação positiva à proteína 16 (p16), e dois à gp 50, de pouca intensidade. Quanto à gp 70, proteína que, embora reagisse com o soro padrão positivo, não foi reconhecida por nenhum dos soros testados. Estes resultados sugerem que o WB pode ser empregado para o sorodiagnóstico rotineiro das lentiviroses, ensejando estudos mais amplos do padrão de reconhecimento dos antígenos apresentados por este novo sistema de purificação parcial de componentes virais.
[Western blot for the diagnosis of small ruminant lentivirus infections in goats: a simple method for antigen production].
Abstract: The serodiagnosis of small ruminant lentivirus (SRLV) infections in goats and sheep is usually performed by the agar-gel immunodiffusion technique (AGID) and by ELISA. Therefore, the western blot (WB) is the choice for the definition of antigen recognition patterns during the disease progression, being potentially useful for the diagnosis itself. In the present study, the antigen used for WB was obtained by a novel simple purification procedure: the dialysis of the supernatant of infected cultured cells, followed by centrifugation in sucrose gradient. Proteins in the pellet were separated by SDS-PAGE at 10% density and transferred to nitrocellulose membranes in a semi-dry blotting device. After development, five viral proteins were recognized by the standard positive serum sample, with molecular weights of 14-16, 25, 40, 50 and 70 kDa respectively. All 8 AGID positive goat serum samples recognized at least one band in the immunoblot, with different intensities. The total number of bands recognized by each serum sample varied considerably. A positive reaction to gp40 was observed with 4 sera, although rather weak in 3 cases. Two samples were reactive to p16 and two others to gp50, although weakly. Curiously, no serum was reactive to the 70 kDa antigen, recognized by the standard positive serum sample. These results suggest that the WB can be effectively used for the routine serodiagnosis of viral caprine arthritis encephalitis infection (CAEV). It can also support further studies on antigen recognition patterns with the new antigen provided by the novel simplified purification system of viral components described here. MenosResumo: O sorodiagnóstico das lentiviroses de caprinos e ovinos é realizado principalmente pela imunodifusão em gel de agar (AGID) e/ou ELISA. Embora relativamente simples, esses métodos não identificam os antígenos virais reconhecidos na resposta imune do animal examinado, por isso o western blot (WB) vem ganhando maior relevância como ferramenta de diagnóstico dessas enfermidades. Neste trabalho, o antígeno utilizado no WB foi obtido através de um sistema simplificado de purificação: concentração por diálise do sobrenadante de culturas celulares infectadas, seguido de centrifugação em gradiente contínuo de sacarose. A separação das proteínas virais foi obtida por SDS-PAGE a 10% e a transferência para membranas de nitrocelulose realizada pelo sistema semi-úmido. A revelação das membranas mostrou reconhecimento pelo soro padrão positivo de cinco proteínas, com pesos moleculares de 14-16, 25, 40, 50 e 70 kDa. Todas as 8 amostras de soro caprino, positivas na ADIG, reconheceram pelo menos uma banda proteíca no immunoblot, variando contudo o número de bandas reconhecidas. Reação positiva à glícoproteína 40 (gp 40) foi observada em quatro animais, com intensidade de reação discreta em três deles. Dois animais apresentaram reação positiva à proteína 16 (p16), e dois à gp 50, de pouca intensidade. Quanto à gp 70, proteína que, embora reagisse com o soro padrão positivo, não foi reconhecida por nenhum dos soros testados. Estes resultados sugerem que o WB pode ser empregado para o s... Mostrar Tudo |
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Palavras-Chave: |
Artrite encefalite caprina; CAEV; Immunoblot; Lentivirose; SRLV; Western blot. |
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Thesagro: |
Antígeno; Caprino; Diagnóstico; Doença animal; Ovino. |
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Thesaurus Nal: |
Lentivirus. |
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Categoria do assunto: |
-- |
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Marc: |
LEADER 04469naa a2200313 a 4500
001 1960451
005 2016-06-27
008 2013 bl uuuu u00u1 u #d
100 1 $aOLIVEIRA, M. M. M.
245 $aWestern blot para o diagnóstico das infecções pelos lentivírus de pequenos ruminantes em caprinos$bum método simples para a produção de antígeno.$h[electronic resource]
260 $c2013
520 $aResumo: O sorodiagnóstico das lentiviroses de caprinos e ovinos é realizado principalmente pela imunodifusão em gel de agar (AGID) e/ou ELISA. Embora relativamente simples, esses métodos não identificam os antígenos virais reconhecidos na resposta imune do animal examinado, por isso o western blot (WB) vem ganhando maior relevância como ferramenta de diagnóstico dessas enfermidades. Neste trabalho, o antígeno utilizado no WB foi obtido através de um sistema simplificado de purificação: concentração por diálise do sobrenadante de culturas celulares infectadas, seguido de centrifugação em gradiente contínuo de sacarose. A separação das proteínas virais foi obtida por SDS-PAGE a 10% e a transferência para membranas de nitrocelulose realizada pelo sistema semi-úmido. A revelação das membranas mostrou reconhecimento pelo soro padrão positivo de cinco proteínas, com pesos moleculares de 14-16, 25, 40, 50 e 70 kDa. Todas as 8 amostras de soro caprino, positivas na ADIG, reconheceram pelo menos uma banda proteíca no immunoblot, variando contudo o número de bandas reconhecidas. Reação positiva à glícoproteína 40 (gp 40) foi observada em quatro animais, com intensidade de reação discreta em três deles. Dois animais apresentaram reação positiva à proteína 16 (p16), e dois à gp 50, de pouca intensidade. Quanto à gp 70, proteína que, embora reagisse com o soro padrão positivo, não foi reconhecida por nenhum dos soros testados. Estes resultados sugerem que o WB pode ser empregado para o sorodiagnóstico rotineiro das lentiviroses, ensejando estudos mais amplos do padrão de reconhecimento dos antígenos apresentados por este novo sistema de purificação parcial de componentes virais. [Western blot for the diagnosis of small ruminant lentivirus infections in goats: a simple method for antigen production]. Abstract: The serodiagnosis of small ruminant lentivirus (SRLV) infections in goats and sheep is usually performed by the agar-gel immunodiffusion technique (AGID) and by ELISA. Therefore, the western blot (WB) is the choice for the definition of antigen recognition patterns during the disease progression, being potentially useful for the diagnosis itself. In the present study, the antigen used for WB was obtained by a novel simple purification procedure: the dialysis of the supernatant of infected cultured cells, followed by centrifugation in sucrose gradient. Proteins in the pellet were separated by SDS-PAGE at 10% density and transferred to nitrocellulose membranes in a semi-dry blotting device. After development, five viral proteins were recognized by the standard positive serum sample, with molecular weights of 14-16, 25, 40, 50 and 70 kDa respectively. All 8 AGID positive goat serum samples recognized at least one band in the immunoblot, with different intensities. The total number of bands recognized by each serum sample varied considerably. A positive reaction to gp40 was observed with 4 sera, although rather weak in 3 cases. Two samples were reactive to p16 and two others to gp50, although weakly. Curiously, no serum was reactive to the 70 kDa antigen, recognized by the standard positive serum sample. These results suggest that the WB can be effectively used for the routine serodiagnosis of viral caprine arthritis encephalitis infection (CAEV). It can also support further studies on antigen recognition patterns with the new antigen provided by the novel simplified purification system of viral components described here.
650 $aLentivirus
650 $aAntígeno
650 $aCaprino
650 $aDiagnóstico
650 $aDoença animal
650 $aOvino
653 $aArtrite encefalite caprina
653 $aCAEV
653 $aImmunoblot
653 $aLentivirose
653 $aSRLV
653 $aWestern blot
700 1 $aMELO, M. A. de
700 1 $aANDRADE, P. P. de
700 1 $aGOMES, S. M.
700 1 $aCAMPOS, A. C.
773 $tArquivo Brasileiro de Medicina Veterinária e Zootecnia$gv. 65, n. 3, p. 694-698, 2013.
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Registro original: |
Embrapa Caprinos e Ovinos (CNPC) |
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Registro Completo
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Biblioteca(s): |
Embrapa Gado de Leite. |
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Data corrente: |
28/11/2018 |
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Data da última atualização: |
24/01/2023 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Circulação/Nível: |
A - 1 |
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Autoria: |
PERIPOLLI, E.; STAFUZZA, N. B.; MUNARI, D. P.; LIMA, A. L. F.; IRGANG, R.; MACHADO, M. A.; PANETTO, J. C. do C.; VENTURA, R. V.; BALDI, F.; SILVA, M. V. G. B. |
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Afiliação: |
ELISA PERIPOLLI, UNESP; NEDENIA BONVINO STAFUZZA, UNESP; DANÍSIO PRADO MUNARI, UNESP / CNPQ; ANDRÉ LUÍS FERREIRA LIMA, UFSC; RENATO IRGANG, UFSC; MARCO ANTONIO MACHADO, CNPGL; JOAO CLAUDIO DO CARMO PANETTO, CNPGL; RICARDO VIEIRA VENTURA, USP / Beef Improvement Opportunities, Canada / University of Guelph, Canada; FERNANDO BALDI, UNESP; MARCOS VINICIUS GUALBERTO B SILVA, CNPGL. |
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Título: |
Assessment of runs of homozygosity islands and estimates of genomic inbreeding in Gyr (Bos indicus) dairy cattle. |
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Ano de publicação: |
2018 |
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Fonte/Imprenta: |
BMC Genomics, v. 19, n. 34, 2018. |
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Páginas: |
13 p. |
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DOI: |
10.1186/s12864-017-4365-3 |
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Idioma: |
Inglês |
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Conteúdo: |
Abstract BACKGROUND: Runs of homozygosity (ROH) are continuous homozygous segments of the DNA sequence. They have been applied to quantify individual autozygosity and used as a potential inbreeding measure in livestock species. The aim of the present study was (i) to investigate genome-wide autozygosity to identify and characterize ROH patterns in Gyr dairy cattle genome; (ii) identify ROH islands for gene content and enrichment in segments shared by more than 50% of the samples, and (iii) compare estimates of molecular inbreeding calculated from ROH (FROH), genomic relationship matrix approach (FGRM) and based on the observed versus expected number of homozygous genotypes (FHOM), and from pedigree-based coefficient (FPED). RESULTS: ROH were identified in all animals, with an average number of 55.12 ± 10.37 segments and a mean length of 3.17 Mb. Short segments (ROH1-2 Mb) were abundant through the genomes, which accounted for 60% of all segments identified, even though the proportion of the genome covered by them was relatively small. The findings obtained in this study suggest that on average 7.01% (175.28 Mb) of the genome of this population is autozygous. Overlapping ROH were evident across the genomes and 14 regions were identified with ROH frequencies exceeding 50% of the whole population. Genes associated with lactation (TRAPPC9), milk yield and composition (IRS2 and ANG), and heat adaptation (HSF1, HSPB1, and HSPE1), were identified. Inbreeding coefficients were estimated through the application of FROH, FGRM, FHOM, and FPED approaches. FPED estimates ranged from 0.00 to 0.327 and FROH from 0.001 to 0.201. Low to moderate correlations were observed between FPED-FROH and FGRM-FROH, with values ranging from -0.11 to 0.51. Low to high correlations were observed between FROH-FHOM and moderate between FPED-FHOM and FGRM-FHOM. Correlations between FROH from different lengths and FPED gradually increased with ROH length. CONCLUSIONS: Genes inside ROH islands suggest a strong selection for dairy traits and enrichment for Gyr cattle environmental adaptation. Furthermore, low FPED-FROH correlations for small segments indicate that FPED estimates are not the most suitable method to capture ancient inbreeding. The existence of a moderate correlation between larger ROH indicates that FROH can be used as an alternative to inbreeding estimates in the absence of pedigree records. MenosAbstract BACKGROUND: Runs of homozygosity (ROH) are continuous homozygous segments of the DNA sequence. They have been applied to quantify individual autozygosity and used as a potential inbreeding measure in livestock species. The aim of the present study was (i) to investigate genome-wide autozygosity to identify and characterize ROH patterns in Gyr dairy cattle genome; (ii) identify ROH islands for gene content and enrichment in segments shared by more than 50% of the samples, and (iii) compare estimates of molecular inbreeding calculated from ROH (FROH), genomic relationship matrix approach (FGRM) and based on the observed versus expected number of homozygous genotypes (FHOM), and from pedigree-based coefficient (FPED). RESULTS: ROH were identified in all animals, with an average number of 55.12 ± 10.37 segments and a mean length of 3.17 Mb. Short segments (ROH1-2 Mb) were abundant through the genomes, which accounted for 60% of all segments identified, even though the proportion of the genome covered by them was relatively small. The findings obtained in this study suggest that on average 7.01% (175.28 Mb) of the genome of this population is autozygous. Overlapping ROH were evident across the genomes and 14 regions were identified with ROH frequencies exceeding 50% of the whole population. Genes associated with lactation (TRAPPC9), milk yield and composition (IRS2 and ANG), and heat adaptation (HSF1, HSPB1, and HSPE1), were identified. Inbreeding coefficient... Mostrar Tudo |
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Palavras-Chave: |
Dairy traits; Inbreeding coefficients; ROH islands. |
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Thesagro: |
Bos Indicus. |
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Categoria do assunto: |
L Ciência Animal e Produtos de Origem Animal |
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URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/item/187250/1/Cnpgl-2018-BMC-Gen-Machado-Assessment.pdf
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Marc: |
LEADER 03293naa a2200301 a 4500
001 2100275
005 2023-01-24
008 2018 bl uuuu u00u1 u #d
024 7 $a10.1186/s12864-017-4365-3$2DOI
100 1 $aPERIPOLLI, E.
245 $aAssessment of runs of homozygosity islands and estimates of genomic inbreeding in Gyr (Bos indicus) dairy cattle.$h[electronic resource]
260 $c2018
300 $a13 p.
520 $aAbstract BACKGROUND: Runs of homozygosity (ROH) are continuous homozygous segments of the DNA sequence. They have been applied to quantify individual autozygosity and used as a potential inbreeding measure in livestock species. The aim of the present study was (i) to investigate genome-wide autozygosity to identify and characterize ROH patterns in Gyr dairy cattle genome; (ii) identify ROH islands for gene content and enrichment in segments shared by more than 50% of the samples, and (iii) compare estimates of molecular inbreeding calculated from ROH (FROH), genomic relationship matrix approach (FGRM) and based on the observed versus expected number of homozygous genotypes (FHOM), and from pedigree-based coefficient (FPED). RESULTS: ROH were identified in all animals, with an average number of 55.12 ± 10.37 segments and a mean length of 3.17 Mb. Short segments (ROH1-2 Mb) were abundant through the genomes, which accounted for 60% of all segments identified, even though the proportion of the genome covered by them was relatively small. The findings obtained in this study suggest that on average 7.01% (175.28 Mb) of the genome of this population is autozygous. Overlapping ROH were evident across the genomes and 14 regions were identified with ROH frequencies exceeding 50% of the whole population. Genes associated with lactation (TRAPPC9), milk yield and composition (IRS2 and ANG), and heat adaptation (HSF1, HSPB1, and HSPE1), were identified. Inbreeding coefficients were estimated through the application of FROH, FGRM, FHOM, and FPED approaches. FPED estimates ranged from 0.00 to 0.327 and FROH from 0.001 to 0.201. Low to moderate correlations were observed between FPED-FROH and FGRM-FROH, with values ranging from -0.11 to 0.51. Low to high correlations were observed between FROH-FHOM and moderate between FPED-FHOM and FGRM-FHOM. Correlations between FROH from different lengths and FPED gradually increased with ROH length. CONCLUSIONS: Genes inside ROH islands suggest a strong selection for dairy traits and enrichment for Gyr cattle environmental adaptation. Furthermore, low FPED-FROH correlations for small segments indicate that FPED estimates are not the most suitable method to capture ancient inbreeding. The existence of a moderate correlation between larger ROH indicates that FROH can be used as an alternative to inbreeding estimates in the absence of pedigree records.
650 $aBos Indicus
653 $aDairy traits
653 $aInbreeding coefficients
653 $aROH islands
700 1 $aSTAFUZZA, N. B.
700 1 $aMUNARI, D. P.
700 1 $aLIMA, A. L. F.
700 1 $aIRGANG, R.
700 1 $aMACHADO, M. A.
700 1 $aPANETTO, J. C. do C.
700 1 $aVENTURA, R. V.
700 1 $aBALDI, F.
700 1 $aSILVA, M. V. G. B.
773 $tBMC Genomics$gv. 19, n. 34, 2018.
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Embrapa Gado de Leite (CNPGL) |
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