RESUMO
Subgroup J avian leukosis virus (ALV-J), first isolated in 1989, mainly induces tumours of myeloid leukosis (ML) in meat-type chickens. In 2006, ALV-J strain SCAU-HN06 was isolated from commercial layer hens with spontaneous haemangiomas in China. To confirm its role in the induction of haemangioma, we constructed a full-length copy of the proviral genome from SCAU-HN06, recovered virus from DF-1 cells detected by enzyme-linked immunosorbent assay, characterized its growth property and investigated its pathogenicity. The recovered virus appeared to be identical to SCAU-HN06 analysed by both blast gene sequences and indirect immunofluorescence assay. It also showed similarities in growth to the parental wild-type virus in vitro. The pathogenicity of the rescued and parental virus in specific-pathogen-free White Leghorn chickens was investigated. Both SCAU-HN06 and rSCAU-HN06 could induce haemangioma, with incidence of 52% and 42.8% respectively. Overall, our findings indicated that the ALV-J strain SCAU-HN06 was the causal agent inducing haemangiomas rather than ML in certain layer chickens.
Assuntos
Vírus da Leucose Aviária/isolamento & purificação , Galinhas , Hemangioma/veterinária , Doenças das Aves Domésticas/virologia , Animais , Vírus da Leucose Aviária/classificação , China/epidemiologia , Feminino , Genoma Viral , Hemangioma/epidemiologia , Hemangioma/patologia , Hemangioma/virologia , Doenças das Aves Domésticas/epidemiologia , Doenças das Aves Domésticas/patologiaRESUMO
In 2005, an avian influenza virus stain was isolated from Parrot in Guangdong, which was then genotyped as H5N2 subtype and designated as A/Parrot/Guangdong/268/2005. According to the current OIE definition on the low-pathogenicity of avian influenza virus, the strain was recognized as a low pathogenic avian influenza virus due to the presence of one basic amino acid residue at the HA cleavage site. Some molecular characteristics of the virus, such as potential glycosylation sites in HA and NA, receptor binding sites of HA, and drug resistance site of NA, showed no variations. To analyze molecular evolution of this strain, we selected the sequences of H5N2 subtype AIVs from GenBank and established the phylogenetic trees. Our results indicated that this strain shared the highest homologies with the H5N2 LPAI isolate A/Pheasant/NJ/1355/1998-like. Phylogenic analysis revealed the isolate, together with A/Chicken/Pennsylvania/1/1983 (H5N2), belonged to America lineages and clustered with A/Pheasant/NJ/1355/1998-like.
Assuntos
Vírus da Influenza A Subtipo H5N2/genética , Vírus da Influenza A Subtipo H5N2/isolamento & purificação , Influenza Aviária/virologia , Papagaios/virologia , Sequência de Aminoácidos , Animais , Evolução Molecular , Genes Virais/genética , Filogenia , Análise de Sequência de DNA , Proteínas Virais/química , Proteínas Virais/genéticaRESUMO
OBJECTIVE: We developed an indirect enzyme-linked immunosorbent assay (ELISA) by the recombinant VP1 protein of duck hepatitis virus (DHV) expressed in Escherichia coli to detect antibodies against DHV. METHODS: The VP1 gene of DHV was amplified by reverse transcription-polymerase chain reaction (RT-PCR) and cloned into pMD18-T and pET-32a vectors to get a prokaryotic expression vector pET-32a-VP1. DHV VP1 gene was expressed and analyzed. A method of enzyme-linked immunosorbent assay (ELISA) was developed and studied. RESULTS: We obtained the recombinant plasmid pET-32a-VP1 with correct sequence and orientation. DHV VP1 gene was expressed in E. coli BL21 (DE3) at a high level and had good immunoreactivity by SDS-PAGE and western blot. The optimum working concentration of antigen was 5.0 microg in 100 microL per well, the working concentration of serum samples was 1 : 100 dilution and the critical value was OD450 > or = 0.302 for the ELISA assay. The rate of coincidence of ELISA and virus neutralization test (VN) was 97.5% by detecting 80 serum samples. The method was specific, sensitive and could be applied for antibody detection of maternal antibody and the rule of antibody growth and decline in immunizing ducklings. CONCLUSION: The ELISA method developed by the purified recombinant protein could be used to detect antibodies against DHV.
Assuntos
Anticorpos/análise , Anticorpos/imunologia , Vírus da Hepatite do Pato/genética , Vírus da Hepatite do Pato/imunologia , Proteínas Virais/genética , Animais , Clonagem Molecular , Patos/crescimento & desenvolvimento , Patos/imunologia , Ensaio de Imunoadsorção Enzimática , Escherichia coli/genética , Feminino , Expressão Gênica , Imunização , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Proteínas Virais/biossíntese , Proteínas Virais/imunologia , Proteínas Virais/isolamento & purificaçãoRESUMO
Coronavirus infection was investigated in apparently healthy wild peafowls in Guangdong province of China in 2003, while severe acute respiratory syndrome (SARS) broke out there. No SARS-like coronavirus had been isolated but a novel avian coronavirus strain, Peafowl/GD/KQ6/2003 (KQ6), was identified. Sequence analysis revealed that KQ6 was an avian coronavirus infectious bronchitis virus (IBV), a member of coronavirus in group 3. The genome sequence of KQ6 had extremely high degree of identity with that of a Massachusetts prototype IBV M41. KQ6 was pathogenic to chickens but non-pathogenic to peafowls under experimental conditions. Seventeen out of fifty-four (31.48%) peafowl serum samples were tested positive for specific antibodies against IBV. Present results indicate that the peafowl isolate KQ6 is a Massachusetts prototype like coronavirus strain which undergoes few genetic changes and peafowl might have acted as a natural reservoir of IBV for very long time.
