| Partially purified homogenates prepared from the livers of birds affectedwith Big Liver and Spleen Disease were used to inoculate intravenously adult broiler breeder hens. After inoculation, the broiler breeders developed pathological changes as same as reported, meanwhile, the serological responses of birds had the positive results of BLS infection. It suggested that the artificial infection of BLS in birds in Nanjing was successful.The gene of big liver and spleen disease virus (BLSV) was amplified from livers of inoculated AA chickens by retrotranscription polymerase chain reaction (RT-PCR). RT-PCR product was cloned into pMD18-T vector and sequenced. The tested sequence shared 97.5% and 84% nt homologue indentities with the reported sequence of BLSV and avian Hepatitis E virus respectively.A RT-PCR assay for the rapid and sensitive detection of BLSV gene was developed. Because of the highly sensitivity for the virus detection, the method could be used for fast diagnosis and epidemic survey of BLSV. BLSV-NJ2002 has been detected by use of RT-PCR in Nanjing. RT-PCR product was cloned into pMD18-T vector and sequenced. Phylogentic tree analysis suggests that the sequence shared 99.5% and 97.3% nt hemologue indentities with BLS V-SD1995 and that of Australia strain respectively. |
