Genome analysis and pathogenicity of reticuloendotheliosis virus isolated from a contaminated vaccine seed against infectious bursal disease virus: first report in China.

Abstract

Specific-pathogen-free (SPF) chickens were inoculated with the virus seed of an infectious bursal disease virus (IBDV)-attenuated vaccine, and positive reticuloendotheliosis virus (REV) antibody levels were subsequently detected in the chicken sera, indicating potential REV contamination of the vaccine. After neutralization with IBDV-positive blood serum, the vaccine was inoculated into DF-1 cells for REV isolation and identification. An REV strain, designated IBD-C1605, was identified using an immunofluorescence assay test. Three pairs of primers were employed for the amplification, cloning and sequencing of three overlapping fragments of the IBD-C1605 genome, and the whole-genome sequence of this isolate was obtained after gene assembly. The genome was 8362 base pairs (nt) in length and its homology with the nucleotide sequences of different reference strains varied between 94.2 and 99.2 %. Isolate IBD-C1605 was inoculated into 1-day-old SPF chickens to observe its pathogenicity. Infection with this organism slowed down the weight gain of SPF chickens and caused atrophy of their immune organs, such as the bursa of Fabricius and thymus gland. Furthermore, the chicken antibody levels decreased significantly after Newcastle disease virus and avian influenza virus subtype H9 vaccine immunization. This is the first report on the isolation and identification of REV from attenuated vaccine virus seeds in China, and is also the first study on the pathogenicity of REV from a contaminated vaccine in China. Our findings contribute towards a better understanding of the detrimental effects of vaccine contamination with exogenous viruses such as REV.