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Registro Completo |
Biblioteca(s): |
Embrapa Unidades Centrais. |
Data corrente: |
15/02/2017 |
Data da última atualização: |
29/12/2017 |
Autoria: |
CRUZ, T. F.; KANASHIRO, T. M.; CASTRO, A. M. M. G. de; BALDIN, C. M.; RICHTZENHAIN, L. J.; ARAUJO JUNIOR, J. P. |
Afiliação: |
TAÍS F. CRUZ, IBTEC/UNESP; TATIANA M. KANASHIRO, DMI/IB/UNESP; ALESSANDRA M. M G. DE CASTRO, FMVZ/USP; CINTIA M. BALDIN, FMVZ/USP; LEONARDO J. RICHTZENHAIN, MVZ/USP; JOÃO PAULO ARAUJO JR, UNESP/IBTEC. |
Título: |
A double-antibody sandwich ELISA based on the porcine circovirus type 2 (PCV2) propagated in cell culture for antibody detection. |
Ano de publicação: |
2016 |
Fonte/Imprenta: |
Pesquisa Veterinária Brasileira, Rio de Janeiro, v. 36, n. 12, p. 1171-1177, dez. 2016. |
Idioma: |
Inglês |
Conteúdo: |
Few studies have described enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies against porcine circovirus type 2 (PCV2) based on antigens produced in cell culture. Furthermore, few articles have described viral purification techniques for members of the family Circoviridae. This occurs because circoviruses are difficult to isolate, noncytopathogenic, and produce low viral titres in cell culture. Thus, for overcoming these difficulties in the cultivation of PCV2, this study aimed to develop a double-antibody sandwich ELISA based on the cell culture antigen PCV2b for the quantification of anti-PCV2 antibodies. A 20% and 50% discontinuous sucrose cushion was used for viral purification, which enabled the separation of cell culture proteins in the 20% sucrose cushion and a greater viral concentration in the 50% sucrose cushion. Following isopycnic centrifugation, PCV2 was concentrated in the band with density values from 1.330 to 1.395g/cm3. Viral purification was assessed using SDS-PAGE, indirect ELISA and electron microscopy. The standardised ELISA revealed a strong linear correlation (r= 0.826, p<0.001) when compared with a commercial ELISA kit. The assay exhibited low variability (inter-assay coefficient of variation of 4.24% and intra-assay of 1.80%) and excellent analytical specificity conferred by the capture antibody produced in rabbit. Thus, this ELISA is a rapid, specific and convenient method for the detection of antibodies against PCV2 in studies of experimental and natural infection, and in monitoring the response to vaccination on commercial farms. MenosFew studies have described enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies against porcine circovirus type 2 (PCV2) based on antigens produced in cell culture. Furthermore, few articles have described viral purification techniques for members of the family Circoviridae. This occurs because circoviruses are difficult to isolate, noncytopathogenic, and produce low viral titres in cell culture. Thus, for overcoming these difficulties in the cultivation of PCV2, this study aimed to develop a double-antibody sandwich ELISA based on the cell culture antigen PCV2b for the quantification of anti-PCV2 antibodies. A 20% and 50% discontinuous sucrose cushion was used for viral purification, which enabled the separation of cell culture proteins in the 20% sucrose cushion and a greater viral concentration in the 50% sucrose cushion. Following isopycnic centrifugation, PCV2 was concentrated in the band with density values from 1.330 to 1.395g/cm3. Viral purification was assessed using SDS-PAGE, indirect ELISA and electron microscopy. The standardised ELISA revealed a strong linear correlation (r= 0.826, p<0.001) when compared with a commercial ELISA kit. The assay exhibited low variability (inter-assay coefficient of variation of 4.24% and intra-assay of 1.80%) and excellent analytical specificity conferred by the capture antibody produced in rabbit. Thus, this ELISA is a rapid, specific and convenient method for the detection of antibodies against PCV2 in stud... Mostrar Tudo |
Palavras-Chave: |
Isopycnic centrifugation; Sucrose cushion. |
Thesaurus Nal: |
Antibody detection; enzyme-linked immunosorbent assay; Porcine circovirus-2. |
Categoria do assunto: |
-- |
URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/item/155768/1/A-double-antibody-sandwich-ELISA.pdf
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Marc: |
LEADER 02442naa a2200241 a 4500 001 2064305 005 2017-12-29 008 2016 bl uuuu u00u1 u #d 100 1 $aCRUZ, T. F. 245 $aA double-antibody sandwich ELISA based on the porcine circovirus type 2 (PCV2) propagated in cell culture for antibody detection.$h[electronic resource] 260 $c2016 520 $aFew studies have described enzyme-linked immunosorbent assays (ELISAs) for the detection of antibodies against porcine circovirus type 2 (PCV2) based on antigens produced in cell culture. Furthermore, few articles have described viral purification techniques for members of the family Circoviridae. This occurs because circoviruses are difficult to isolate, noncytopathogenic, and produce low viral titres in cell culture. Thus, for overcoming these difficulties in the cultivation of PCV2, this study aimed to develop a double-antibody sandwich ELISA based on the cell culture antigen PCV2b for the quantification of anti-PCV2 antibodies. A 20% and 50% discontinuous sucrose cushion was used for viral purification, which enabled the separation of cell culture proteins in the 20% sucrose cushion and a greater viral concentration in the 50% sucrose cushion. Following isopycnic centrifugation, PCV2 was concentrated in the band with density values from 1.330 to 1.395g/cm3. Viral purification was assessed using SDS-PAGE, indirect ELISA and electron microscopy. The standardised ELISA revealed a strong linear correlation (r= 0.826, p<0.001) when compared with a commercial ELISA kit. The assay exhibited low variability (inter-assay coefficient of variation of 4.24% and intra-assay of 1.80%) and excellent analytical specificity conferred by the capture antibody produced in rabbit. Thus, this ELISA is a rapid, specific and convenient method for the detection of antibodies against PCV2 in studies of experimental and natural infection, and in monitoring the response to vaccination on commercial farms. 650 $aAntibody detection 650 $aenzyme-linked immunosorbent assay 650 $aPorcine circovirus-2 653 $aIsopycnic centrifugation 653 $aSucrose cushion 700 1 $aKANASHIRO, T. M. 700 1 $aCASTRO, A. M. M. G. de 700 1 $aBALDIN, C. M. 700 1 $aRICHTZENHAIN, L. J. 700 1 $aARAUJO JUNIOR, J. P. 773 $tPesquisa Veterinária Brasileira, Rio de Janeiro$gv. 36, n. 12, p. 1171-1177, dez. 2016.
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Embrapa Unidades Centrais (AI-SEDE) |
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Registros recuperados : 5 | |
1. | | FAVERO, C. M.; PIOVEZAN, U.; CASTRO, A. M. M. G; SZABO, M.; HERRERA, H. M.; SILVA, S.O.S; BRANDÃO, P. E.; RICHTZENHAIN, L. J. Caracterização genética de torque teno virus, genótipo 1 e 2, detectados em soro de porco monteiro (Sus Scrofa) do Pantanal, Mato Grosso do Sul. In: CONGRESSO DA ABRAVES, 15., 2011, Fortaleza. [Anais...]. Fortaleza: ABRAVES, 2011. Não paginado. ASSOCIAÇÃO BRASILEIRA DOS VETERINÁRIOS ESPECIALISTAS EM SUÍNOS.Tipo: Artigo em Anais de Congresso |
Biblioteca(s): Embrapa Pantanal. |
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2. | | CRUZ, T. F.; KANASHIRO, T. M.; CASTRO, A. M. M. G. de; BALDIN, C. M.; RICHTZENHAIN, L. J.; ARAUJO JUNIOR, J. P. A double-antibody sandwich ELISA based on the porcine circovirus type 2 (PCV2) propagated in cell culture for antibody detection. Pesquisa Veterinária Brasileira, Rio de Janeiro, v. 36, n. 12, p. 1171-1177, dez. 2016.Biblioteca(s): Embrapa Unidades Centrais. |
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3. | | BALDIN, C. M.; FAVERO, C. M.; CASTRO, A. M. M. G.; PIOVEZAN, U.; JULIANO, R. S.; HERRERA, H. M.; SZABO, M.; BRANDÃO, P. E.; RICHTZENHAIN, L. J. Detecção de Porcine circovirus2 (PCV2) e Torque teno sus virus (TTsuV1 e TTSuV2) e amostras de soro de porco-monteiro do Pantanal Sul-Matogrossense. In: CONGRESSO DA ABRAVES, 15., 2011, Fortaleza. [Anais...]. Fortaleza: ABRAVES, 2011. Não paginado. ASSOCIAÇÃO BRASILEIRA DOS VETERINÁRIOS ESPECIALISTAS EM SUÍNOS.Tipo: Artigo em Anais de Congresso |
Biblioteca(s): Embrapa Pantanal. |
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4. | | FRANZO, G.; CORTEY, M.; CASTRO, A. M. M. G. de; PIOVEZAN, U.; SZABO, M. P. J.; DRIGO, M.; SEGALÉS, J.; RICHTZENHAIN, L. J. Genetic characterisation of Porcine circovirus type 2 (PCV2) strains from feral pigs in the Brazilian Pantanal: An opportunity to reconstruct the history of PCV2 evolution. Veterinary Microbiology, v. 178, n. 1-2, p. 158-162, 2015.Tipo: Artigo em Periódico Indexado | Circulação/Nível: A - 1 |
Biblioteca(s): Embrapa Pantanal. |
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5. | | CASTRO, A. M. M. G; FERRARI, K. L.; BALDIN, C. M.; CASTRO JUNIOR, E. G.; PIOVEZAN, U.; JULIANO, R. S.; HERRERA, H. M.; SZABO, H. M.; BRANDÃO, P. E.; RITCHTZENBAIN, L. J. Genetic characterization and phylogenetic analysis of porcine circovirus 2 (PCV2) detected in diferent susceptible Sus Scrofa. Virus: reviews and research, v.16, sup. 1, p. 294, oct. 2011. Resumo apresentado no XXII National Meeting of Virology & VI mercosur Meeting of Virology. VV510.Tipo: Resumo em Anais de Congresso |
Biblioteca(s): Embrapa Pantanal. |
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Registros recuperados : 5 | |
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