Sandwich ELISA com duplo anticorpo baseado no circovirus suíno tipo 2 (PCV2) produzido em cultivo celular para detecção de anticorpos
| Autor: | João Pessoa Araújo, Alessandra Marnie Martins Gomes de Castro, Cíntia M Baldin, Leonardo José Richtzenhain, Tatiana M. Kanashiro, Taís F. Cruz |
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| Přispěvatelé: | Universidade Estadual Paulista (Unesp), Universidade de São Paulo (USP) |
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
centrifugação isopícnica 040301 veterinary sciences Coefficient of variation animal diseases Double antibody sandwich isopycnic centrifugation sucrose cushion 0403 veterinary science 03 medical and health sciences Antigen Enzyme-linked immunosorbent assay circovirus suíno tipo 2 anticorpo Antibody Differential centrifugation lcsh:Veterinary medicine General Veterinary biology colchão de sacarose 04 agricultural and veterinary sciences biology.organism_classification Virology Porcine circovirus 030104 developmental biology Cell culture biology.protein lcsh:SF600-1100 VIROSES EM ANIMAIS antibody Antibody detection porcine circovirus type 2 |
| Zdroj: | Pesquisa Veterinária Brasileira, Volume: 36, Issue: 12, Pages: 1171-1177, Published: DEC 2016 Pesquisa Veterinária Brasileira v.36 n.12 2016 Pesquisa Veterinária Brasileira Colégio Brasileiro de Patologia Animal (CBPA) instacron:EMBRAPA Repositório Institucional da USP (Biblioteca Digital da Produção Intelectual) Universidade de São Paulo (USP) instacron:USP Scopus Repositório Institucional da UNESP Universidade Estadual Paulista (UNESP) instacron:UNESP Pesquisa Veterinária Brasileira, Vol 36, Iss 12, Pp 1171-1177 |
| Popis: | 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 |
| Databáze: | OpenAIRE |
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