The Comparation Of Different Laboratory Detection Methods And Research Of Diagnosis Epitope For Monkey B Virus

Objective Monkey B virus(BV),also known as Cercopithecine herpes virus,was significant zoonotic pathogen,and performence should limited in ABSL-4.Alternative antigens were usually used in the detection of B virus from rhesus monkey sera,because of its strict demand for biosafity preparation.But the accuracy of these different detection methods using alternative antigens still need to be verified.Method 135 sera samples from rhesus were detected by HVP2-ELISA、BV-ELISA、HSV-1-IEA and HSV-1-IFA,those candidate indeterminate samples and positive samples were then verified by HSV-1-WB and HSV-1 gCl immunoblotting.Results The B virus positive rate of rhesus monkeys were 36.3%by HVP2-ELISA,38.5%by BV-ELISA,35.6%by HSV-1-IEA and 34.1%by HSV-1-IFA respectively.91.9%(124/135)of the 135 samples tested by the three methods were consistent,and the results of all samples above could be confirmed by WB and IFA.The rest indeterminate 12 samples were further detected by HSV-1-WB and HSV-1 gCl immunoblotting,27.3%(3/11)of samples were confirmed as positive.Conclusion These detection methods,with high coincidence rate,were suitable for screening animals infected with BV.Also,positive samples and suspicious samples need to be confirmed by a variety of methods in case of leak.In addition,in screening tests by ELISA,the criteria of positive samples could be appropriately reduced in case of leak of suspicious samples.Objective To identify B virus specific detecting antigen epitope.Methods Analyzed and compared the whole virus sequence of BV and HSV by bioinformatics prediction technology,and selected the specific and possible linear epitope of B virus for peptide synthesis;At the same time,using the overlap technology,synthesized overlap peptide according to the amino acid sequence of gB and gD,the core antigen protein of B virus.By peptide ELISA,the standard serums samples with B virus antibody evaluated the specificity and sensitivity of peptides.Results the results of pep-ELISA determination show that the antibody of monkey B virus could be detected by the peptide of gD(sequence AQLPPNWHVPEAS)and the peptide of gB(sequence YVRELLREQERRPGDAAATPKPSA),compared with the results of the traditional ELISA,the sensitivity was 96%(48/50)by the combination of two detection peptide;However,the predicted specific epitopes of B virus by bioinformatics could not be proven in experiments for many times.Conclusion the peptide antigens,compared with the antigens of virus particles,had a relatively consistent accuracy and high sensitivety,so this kind of antign could be used as detection antigen for monkey B virus;the predicted specific epitopes of B virus by bioinformatics located on function genes of B virus,and revealed low antigencity,however the epitopes with diagnosis application value still located in the core antigen proteins of B virus.