3 Diagnostic methods of Foot-and-Mouth Disease using recombinant 3ABC non-structural protein expressed in insect cells and monoclonal antibody

摘要

PURPOSE: Diagnostic methods of foot-and-mouth disease using recombinant 3ABC non-structural protein expressed in insect cells and a monoclonal antibody is provided, thereby more rapidly and accurately diagnosing the foot-and-mouth disease than the prior methods. CONSTITUTION: A gene encoding a recombinant 3ABC non-structural protein derived from Korean foot-and-mouth disease virus O/SKR/2000 has the nucleotide sequence of SEQ ID NO: 1. A recombinant baculovirus to be expressed in insect cells is prepared by co-transfection with a recombinant vector containing the recombinant 3ABC non-structural protein gene of SEQ ID NO: 1. The recombinant 3ABC non-structural protein is expressed by infection of the insect cells with the recombinant baculovirus. A hybridoma cell line 3F-11(KCTC 10138BP) is prepared by introducing the recombinant 3ABC non-structural protein expressed in E. coli into an animal, collecting an immunized cell from the animal and fusing the immunized cell with a cancer cell. A monoclonal antibody is produced from the hybridoma cell line 3F-11(KCTC 10138BP). A diagnostic method of foot-and-mouth disease comprises the steps of: (1) diluting the recombinant 3ABC non-structural protein in a coating buffer solution and pouring the diluate on a plate; (2) reacting the testing serum with the plate; (3) reacting a conjugate, which binds with an antibody for foot-and-mouth disease virus in the testing serum and has an enzyme, a radioactive material or a fluorescent material, with the testing serum; and (4) measuring intensity of the enzyme reaction, fluorescence reaction or radiation reaction with the conjugate.