Enzyme-Immunosorbent Assay for Detection of Porcine Infectious Gastroenteritis Virus Antibody Using Genetically Modified Baculovirus and Recombinant Porcine Infectious Gastritis Virus Spike Protein

摘要

Spike protein, a structural protein of the swine transmissible gastroenteritis virus (TGEV), is known to induce the production of neutralizing antibodies that can neutralize TGEV. After separating and identifying TGEV from the dead pigs, the nucleotide sequence of the spike gene was analyzed, and the recombinant TGEV spike protein was inserted by inserting the TGEV spike gene into the baculovirus gene, which is an insect virus, using the genetic recombination method. It was expressed in the vertical (Sf9). When the expressed recombinant TGEV spike protein was inoculated into guinea pigs, it was confirmed using the serum neutralization test to produce neutralizing antibodies against TGEV. Therefore, the neutralized antibody titer of TGEV can be detected using a recombinant TGEV spike protein by a monoclonal antibody capture enzyme linked immunosorbent assay (MACELISA) using TGEV-specific monoclonal antibodies. Currently, the method of measuring antibody titer against TGEV is a virus neutralization test using tissue culture, which takes about 4-5 days of difficult tissue culture and testing, but the method of MACELISA used in the present invention can be used for the recombination protein reaction step. If kitted, neutralizing antibodies in TGEV can be tested within 3 hours.