Publications by authors named "Verkhovskii O"

The results obtained using the diagnostic kit based on real-time polymerase chain reaction to detect the DNA of the African Swine Fever in the pathological material, as well as in the culture fluid, are presented. A high sensitivity and specificity for detection of the DNA in the organs and tissues of animals was shown to be useful for detection in the European Union referentiality reagent kits for DNA detection by real time PCR of ASFV. More rapid and effective method of DNA extraction using columns mini spin Quick gDNA(TM) MiniPrep was suggested and compared to the method of DNA isolation on the inorganic sorbent.

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African swine fever (ASF) is an infectious viral disease that causes high economic losses due to the necessity of depopulation of pigs in affected areas, sanitary measures, trade restrictions, etc. The virus (ASFV) is relatively stable in the unprocessed meat products and environment. Thus, large areas are at risk due to free movement of people and products.

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A panel of hybridomas producing monoclonal antibodies (MAbs) to nucleocapsid protein (NP) of avian influenza A virus was obtained. On the basis of 2 MAbs, the authors designed an antigen-bound ELISA (sandwich ELISA), in which NP3 MAbs were used as antigen-bound antibodies and NP MAbs conjugated with horse radish peroxidase as antigen detection antibodies. The specificity of the test system to avian influenza virus was determined.

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Recombinant nucleocapsid (rN) protein N of porcine reproductive and respiratory syndrome virus (PRRSV) was prepared, by using the E. coli expressiom system. Insertion of a polyhistidine marker into the structure of the protein allowed the latter to be purified by metal-chelate affinity chromatography.

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Recombinant antigen ORF2 from porcine circovirus type 2 (PCV-2) was produced, by using the baculovirus expression system, with histidine tags to allow purification by metal-chelate affinity chromatography. The purity of the protein was verified by polyacrylamide gel electrophoresis; and its immunospecificity was confirmed by the immunoblotting test using reference PCV-2-positive and PCV-2-negative porcine sera and monoclonal antibodies. The protein was used as an antigen to develop an indirect enzyme immunoassay (EIA) of PCV-2 antibodies.

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Full-length Bos taurus PrPC protein was obtained in the eu- and prokaryotic expression systems. Immunoblotting and indirect enzyme immunoassay demonstrated high specificity and antigenic activity of full-length proteins in the reactions with monoclonal antibodies (anti-SAF-32 and VRQ-84). Membrane location of recombinant PrPC protein in insect cells was shown by immunofluorescent analysis.

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A diagnostic test system based on polymer latex carriers sensitized by IgG to Canicola and Icterohaemorrhagiae serogroup Leptospira was developed and tried.

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