[Construction and recovery of chimeric rabies virus expressing envelop proteins E1E2 of hepatitis C].

Objective: Construction and recovery of chimeric rabies virus expressing HCV envelop proteins E1E2.

Methods: On the basis of the previously established reverse genetic system CTN-GFP, HCV E1E2 genes were cloned to both replication competent and replication constrained viral vectors based on CTN181 strain and the chimeric viruses CTN-HCV E1E2 and CTNdeltaG-HCV E1E2 were recovered.

Results: The result demonstrated that both the chimeric viruses were rescued successfully, had the ability to re-infect normal sensitive cell lines and express HCV E1E2 genes detected in the level of mRNA.

[Expression and characterization of rabies virus nucleoprotein in baculovirus].

To construct a recombinant expression plasmid Bacmid-N containing the N gene of Rabies Virus, the N gene of RV CVS-11 strain was cloned into the baculovirus shuttle vector (Bacmid). Recombinant Baculovirus AcMNPV-N (P1 Viral stock) was obtained by transfecting the Bacmid-N into the insect cell line of Sf9. The expressed nucleoprotein was identified and analysised by ELISA, FA, SDS-PAGE and Western blot assays.

[Construction and analysis of full-length cDNA clone of rabies virus street strain].

To construct a expression plasmid containing the full-length cDNA of rabies virus, four overlapped fragments covering full length cDNA of rabies virus street stain HN10 were cloned into pVAX1 sequentially in the genome except for the G-L noncoding region which was replaced with GFP gene. The plasmid containing the full-length viral cDNA was flanked by hammerhead ribozyme (HamRz) and hepatitis delta ribozyme (HdvRz) sequences and arranged under the control of the cytomegalovirus (CMV) promoter. The constructed plasmid could be directly used for the following procedure of producing the recombinant rabies virus street HN10.

[Characterization of RdRp gene of rabies virus in China].

Objective: Characterization of RdRp gene (L) of rabies virus aG and CTN181 strain in China.

Methods: Overlapped fragments were amplified and assembled. Then characterization and phylogenetic analyses as well as prediction of functional regions were performed using biologic softwares.

[The primary application of direct rapid immunohistochemical test to rabies diagnosis in China].

Objective: Evaluation of the direct rapid immumohistochemical test (DRIT) for laboratory surveillance of rabies.

Methods: 72 brain specimens of domestic dogs or patients collected from Guizhou, Guangxi, Hunan, Anhui, Jiangsu and Yunnan provinces were detected by conventional methods including Direct Fluorescent-antibody Assay (DFA) and Reverse Transcription Polymerase Chain Reaction (RT-PCR), and by DRIT which was newly developed in the Rabies Section of the Centers for Disease Control and Prevention in the United States. The sensitivity and specificity of DRIT were evaluated by compare of the three results.