Comparison of different detection methods of monkey B virus antibody / 中国比较医学杂志

Objective Monkey B virus(BV), also known as Cercopithecine herpesvirus 1,is an important zoonotic pathogen.According to the national standard, antibodies are detected using BV as an antigen.However, the preparation of BV antigen is very stricted due to biosafety issues.Therefore, in this study, we used alternative antigens to detect the BV antibody by serological assay and verified their specifity and sensitivity.Methods A total of 135 blood samples from rhesus monkeys were tested by two ELISA method (BV and HVP2) and enzyme immunosorbent assay (EIA)method.The positive and suspicious samples were verified by immuno-fluorescence assay (IFA), Western blot and immunoblotting technique using HSV-1 gC1 purified glycoprotein as an antigen.Results The positive rates of HVP2-ELISA, BV-ELISA and HSV-1-EIA were 32.6%, 37.8% and 34.8%, respectively.Consistant result of the three detection method accounted for 91.1% (123/135), and the positive result were confirmed by IFA And WB.There were 12 suspicious samples,in which 33.3% (4/12) were verified to be positive.Conclusions Compared with BV antigen, the sensitivity and specificity of the alternative antigen HSV-1 are moe close than HVP2.Positive and suspicious samples should be verified by several method to avoid missed detection.

Construction of eukaryotic vector of monkey B virus glycoprotein D gene and the gD gene expression / 中国比较医学杂志

Objective To establish an eukaryotic vector of monkey B virus glycoprotein D gene and analyze the expression of gD gene in human embryonic kidney 293T cells.Method First, the protein of monkey B virus glycoprotein D was obtained by gene synthesis.The gene fragments were digested with Pst I and Not I, and ligated to pEGPF-N3. Then, the recombinant plasmid pEGPF-N3-GD was transfected into 293T cells.The expression of gD protein in the cells was detected by Western blot, and the expression localization was investigated using laser scanning confocal microscopy. Results The recombinant plasmid pEGPF-N3 carrying gD gene was successfully constructed, and normally expressed in the 293T cells.Conclusions Glycoprotein D of monkey B virus is expressed successfully in the 293T cells and the protein is located on the cell surface.It may be useful for the preparation of specific recombinant antigen to the glycoprotein D of monkey B virus on cell surface, and can be also used for preparation of antigen slide for detection of monkey B virus.

Prediction and identification of B-cell epitopes on monkey B virus envelope protein gD / 中国比较医学杂志

Objective To Identify B-cell epitopes on monkey B virus envelope protein gD.Methods Base on bioinformatics software, secondary structure, hydrophilicity, surface accessibility, antigenic index and flexibility of monkey B virus envelope protein gD was analysed and some potential peptide epitopes were forecasted.Then, the interactions between synthetic peptides and BV positive sera were detected by Enzyme-linked immunosorbent assay ( ELISA) .At last, The sensitivity and specificity of synthetic peptides was evaluated used 20 samples of Standard sera by ELISA.Result Seven epitopes were forecasted by Bioinformatics analysis. Four synthetic peptides, sequence as 46 LPPLEQKTD54 , 106 RGAPEATRSDA116 , 291 PELAPEERGTSRTPGD306 and 361 AVYLVRRRGR370 could be reacted with positive sera pool.The sensitivity of 4 synthetic peptides changed form 40% to 70% and the specificity were 100% for 4 synthetic peptides. Conclusion There are at least four linear epitope on B virus gD protein.