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Registro Completo |
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Biblioteca(s): |
Embrapa Unidades Centrais. |
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Data corrente: |
15/02/2017 |
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Data da última atualização: |
29/12/2017 |
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Autoria: |
CRUZ, T. F.; KANASHIRO, T. M.; CASTRO, A. M. M. G. de; BALDIN, C. M.; RICHTZENHAIN, L. J.; ARAUJO JUNIOR, J. P. |
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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. |
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Título: |
A double-antibody sandwich ELISA based on the porcine circovirus type 2 (PCV2) propagated in cell culture for antibody detection. |
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Ano de publicação: |
2016 |
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Fonte/Imprenta: |
Pesquisa Veterinária Brasileira, Rio de Janeiro, v. 36, n. 12, p. 1171-1177, dez. 2016. |
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Idioma: |
Inglês |
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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 |
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Palavras-Chave: |
Isopycnic centrifugation; Sucrose cushion. |
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Thesaurus Nal: |
Antibody detection; enzyme-linked immunosorbent assay; Porcine circovirus-2. |
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Categoria do assunto: |
-- |
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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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Registro original: |
Embrapa Unidades Centrais (AI-SEDE) |
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 | Acesso ao texto completo restrito à biblioteca da Embrapa Florestas. Para informações adicionais entre em contato com cnpf.biblioteca@embrapa.br. |
Registro Completo
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Biblioteca(s): |
Embrapa Florestas. |
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Data corrente: |
06/05/2023 |
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Data da última atualização: |
08/05/2023 |
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Tipo da produção científica: |
Capítulo em Livro Técnico-Científico |
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Autoria: |
RAMIREZ, C.; ALBERDI, I.; BAHAMONDEZ, C.; FREITAS, J. V. de. |
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Afiliação: |
CARLA RAMIREZ, FAO; ICIAR ALBERDI, INIA, CSIC; CARLOS BAHAMONDEZ, INSTITUTO FLORESTAL DE CHILE; JOBERTO VELOSO DE FREITAS, UNIVERSIDADE FEDERAL DO AMAZONAS. |
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Título: |
National forest inventories of Latin America and the Caribbean. |
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Ano de publicação: |
2021 |
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Fonte/Imprenta: |
Roma: FAO, 2021. |
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Páginas: |
407 p. |
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ISBN: |
978-92-5-136394-2 |
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DOI: |
https://doi.org/10.4060/cb7791en |
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Idioma: |
Espanhol |
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Conteúdo: |
National forest inventories (NFIs) are one of the main sources of forest information. This book describes the importance and history of NFIs in Latin America and the Caribbean, a region that is particularly relevant due to the extension and biodiversity of its forests. Methodologies for data collection and measurement of the most relevant indicators in 21 countries are addressed. In addition, similarities and differences in IFN designs, challenges and opportunities, and prospects for the future are examined. This analysis demonstrates that the information generated by the countries is diverse and must be harmonized to meet the commitments and opportunities for sustainable forest management in the 21st century. This publication represents a milestone in the beginning of the harmonization process towards data transparency within the forestry sector in Latin America and the Caribbean and constitutes the first collaborative effort of a network of NFI experts and collaborators in the region. |
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Palavras-Chave: |
Florestas; Manejo florestal. |
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Thesagro: |
Inventário Florestal. |
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Categoria do assunto: |
K Ciência Florestal e Produtos de Origem Vegetal |
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Marc: |
LEADER 01581nam a2200217 a 4500
001 2153546
005 2023-05-08
008 2021 bl uuuu 00u1 u #d
020 $a978-92-5-136394-2
024 7 $ahttps://doi.org/10.4060/cb7791en$2DOI
100 1 $aRAMIREZ, C.
245 $aNational forest inventories of Latin America and the Caribbean.
260 $aRoma: FAO$c2021
300 $a407 p.
520 $aNational forest inventories (NFIs) are one of the main sources of forest information. This book describes the importance and history of NFIs in Latin America and the Caribbean, a region that is particularly relevant due to the extension and biodiversity of its forests. Methodologies for data collection and measurement of the most relevant indicators in 21 countries are addressed. In addition, similarities and differences in IFN designs, challenges and opportunities, and prospects for the future are examined. This analysis demonstrates that the information generated by the countries is diverse and must be harmonized to meet the commitments and opportunities for sustainable forest management in the 21st century. This publication represents a milestone in the beginning of the harmonization process towards data transparency within the forestry sector in Latin America and the Caribbean and constitutes the first collaborative effort of a network of NFI experts and collaborators in the region.
650 $aInventário Florestal
653 $aFlorestas
653 $aManejo florestal
700 1 $aALBERDI, I.
700 1 $aBAHAMONDEZ, C.
700 1 $aFREITAS, J. V. de
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Embrapa Florestas (CNPF) |
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