| Objectives: Emerging virus disease is a new public health challenge in this century. We always don't know what to do when facing these large-scale outbreak virus infections. But these infections can be controlled and cured if we can know the infection mechanisms in advance, expected the possibility of break out and building up reliable and effective detection systems and treatment measures.Borna Disease Virus (BDV), one of the central nervous system emerging viruses, was first found in a small town of Germany and caused outbreak of encephalitis and death in horses, which made people thought it as a potent virus at first. It only caused latent chronic infection and psychiatric symptoms in human infection. Since the outbreak of BDV has precedents in animals, we cannot rule out the possibility of BDV outbreak in human by natural variability. Researchers even found and published it in Nature that part of BDV P protein gene was integrated into the genome of human and several kinds of animals, which may be the pathogenesis of many diseases of human. Establish reliable BDV detection methods and plan different combinations of detection methods under different needs as well as use these methods in studies of BDV infection mechanisms are of great important.Methods:1. Compare and assessment of BDV P24 and BDV P40 RT PCR detection by sensitivity, specificity, reproducibility, stability, and practical application.2. Establish BDV detection cell model and In situ PCR detection. Compare the detection with antibody detection and application on RT PCR positive samples.3. Detect CIC by ELISA and compare to RT PCR and antibody detection.4. Compare all of these detections and establish different combinations of different situations.5. Observation subcellular localization and expression of phosphoprotein at different periods after transfection by In situ PCR, ELISA and Fluorescence microscope on BDV detection cell model. Explore the influence of phosphoprotein transfection by MTT.Results:1. The sensitivity, specificity, reproducibility, stability, and practical application of these two detection methods are all good and got 3.6% (both P24 and P40) positive in human; 4.4% (P24) and 4.2% (P40) positive in pig; 1.7% (P24) and 1.5% (P40) positive in horse. These detections can be use as reliable tools in BDV epidemiological investigations and laboratory studies.2. In situ PCR detection was established and verificated on OL BDV detection models and successful applied on human samples.3. The positive rate of CIC detection is 36%, which is much more than RNA detection and antibody detection. The results showed there are more BDV carriers than common infection patients.4. Different detection combinations of different situations were established by comparing all of these detections.5. Nucleic acid of BDV Phosphoprotein was found in cell nucleus at early time after infection and all over the cell later. No significant differences between different periods of protein location and expression. BDV Phosphoprotein can hold back growth of OL cell which was confirmed by MTT.Conclusion: Follow the different characteristics of BDV RT PCR, In situ PCR, Antibody detection and ELISA for CIC, different detection combinations for different situations were formed. BDV infection mechanisms were studied by these detection tools. Location of phosphoprotein nucleic acid and protein in OL cell after transfection is the same as ture infection and it inhibited the growth of OL cell.
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