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1.
Chinese Journal of Experimental and Clinical Virology ; (6): 405-408, 2011.
Article in Chinese | WPRIM | ID: wpr-246228

ABSTRACT

<p><b>OBJECTIVE</b>To express St. Louis encephalitis virus-like particles in mammalian cells, it will provide a prerequisite for further immunological diagnostic studies.</p><p><b>METHODS</b>293T-cell were transiently transfected with recombinant PreM-E plasmid. Expression and antigenicity of the purified protein were determined by transmission electron microscope (TEM), Western-Blot, immunofluorescence assay and ELISA.</p><p><b>RESULTS</b>Recombinant subviral particles, about 50 nm in diameter, were observed by TEM in the supernatant of transfected cells. The results of Western-Blot, IFA and ELISA showed the recombinant proteins retained immunoreactivity similar to those of native virus proteins.</p><p><b>CONCLUSION</b>St. Louis encephalitis virus-like particles has good antigenicity and physical appearance. It will provide a prerequisite for further immune diagnostic reagent.</p>


Subject(s)
Humans , Allergy and Immunology , Diagnosis , Enzyme-Linked Immunosorbent Assay , HEK293 Cells , Plasmids , Recombinant Proteins , Allergy and Immunology , Transfection , Virion , Allergy and Immunology
2.
Chinese Journal of Experimental and Clinical Virology ; (6): 340-342, 2010.
Article in Chinese | WPRIM | ID: wpr-316905

ABSTRACT

<p><b>OBJECTIVE</b>Through observing the morphological changes of the prepared influenza viruses (H1N1) treated with the different concentration of Nonidet P-40 solutions and added with antibody against the influenza virus using transmission electron microscopy (TEM), to explore the principle of application of the internal antibody for immunoassay of influenza virus.</p><p><b>METHODS</b>Through treating the virus samples with serial diluted Nonidet P-40 solutions from 0.01% to 0.2% and/or not adding antibody against the influenza virus, then investigating the samples by TEM to obtain the morphological changes of the virions.</p><p><b>RESULTS</b>The serial images show that the denudation degree of the virions is proportional with the rise of NP-40 concentration, and partly denuded virion image appeared at 0.1% NP-40 treatment, also under this condition no influence was observed on antibody binding to the virus.</p><p><b>CONCLUSION</b>This work demonstrated the interaction between influenza virion and its antibody under nonionic surfactants existing, which supports the advantages of sample preparation for immunoassay enveloped virus using internal antibody theoretically.</p>


Subject(s)
Animals , Chick Embryo , Antibodies, Viral , Chemistry , Influenza A Virus, H1N1 Subtype , Microscopy, Electron, Transmission , Polyethylene Glycols , Pharmacology , Protein Binding , Viral Envelope Proteins , Chemistry , Virion
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