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Título: Porcine circovirus 2b: isolations and mutation rate in J744 lineage macrophage cell
Título(s) alternativo(s): Circovírus suíno 2b: isolamento e taxa de mutação em linhagem de célula de macrófagos (J744)
Autor(es): Castro, Alessandra Marnie Martins Gomes de
Bersano, Josete Garcia
Souza, Sheila Oliveira Silva
Ogata, Renato Akio
Nara, Julia Mitico
Richtzenhain, Leonardo José
Data do documento: 3-Ago-2017
Resumo: Porcine circovirus 2 (PCV2) is associated with various clinical signs that are collectively designated as Circovirosis and has a great impact on the pig industry. The virus isolation is classically performed on PK-15 cell line, but other cells have been tested. Despite advances in studies with PCV2, isolation is still a challenge. The difficulty of maintaining these cell lines commonly used associated with the use of toxic substances to the isolation of PCV2 had stimulated the present study, that had the objectives to describe the first isolation of PCV2b in macrophage cell lines, J744 and verify the mutation rate at this system. A sample of lung was pooled and submitted to sequencing in which was classified in genotype PCV2b. This sample was used to inoculate a bottle of J744 with 30% of confluence in RPMI with 10% fetal bovine serum and submitted to five passages, which were accompanied by chain reaction quantitative polymerase (PCRq). The initial and final viral loads were 2.90 × 103 and 4.45 × 108 DNA copies/µL for PCV2b, respectively. Sequencing confirmed the isolation and had eliminated possible co-isolation of more than one genotype. After five passages, the isolate showed 99.7% identity with description of five point of non-synonymous or/and synonymous mutations observed in the cap and rep gene. The results demonstrate that J744 cells exhibit susceptibility, and the instability of the virus in J744 will be important for understanding the virus.
Palavras-chave: Disease
Cell culture
viral evolution
Citação: Castro, Alessandra Marnie Martins Gomes de; Bersano, Josete Garcia; Souza, Sheila Oliveira Silva; Ogata, Renato Akio; Nara, Julia Mitico; Richtzenhain, Leonardo José. Porcine circovirus 2b: isolations and mutation rate in J744 lineage macrophage cel. Arquivos do Instituto Biológico, v. 84, e1012014, 2017.
ISSN: 1808-1657
Número DOI: 10.1590/1808-1657001012014
Idioma: en_US
Editora: Instituto Biológico
Local da Publicação: São Paulo
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