3 A diagnostic method of Foot-and-Mouth Disease using recombinant 3ABC non-structural protein expressed in E.coli and monoclonal antibody

PURPOSE: Diagnostic methods of foot-and-mouth disease using a recombinant 3ABC non-structural protein expressed in E. coli 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. The recombinant 3ABC non-structural protein is expressed from a recombinant E. coli transformed with a recombinant vector containing the recombinant 3ABC non-structural protein gene of SEQ ID NO: 1. 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 dilution on a plate; (2) reacting the testing serum with the plate; (3) reacting the 3ABC non-structural protein specific monoclonal antibody with the serum; (4) reacting a 3ABC non-structural protein specific monoclonal antibody binding conjugate with the monoclonal antibody; and (5) measuring intensity of the enzyme reaction, fluorescence reaction or radiation reaction with the conjugate. 본 발명은 구제역 바이러스(Foot-and-Mouth Disease Virus; FMDV) O/SKR/2000유래의 대장균 발현 재조합 3ABC 비구조단백질(Non-Structural Protein; NSP) 및 3ABC 단백질에 특이적인 단클론 항체를 이용한 구제역의 진단방법에 관한 것이다. 더욱 상세하게는, 본 발명은 구제역 바이러스의 3ABC 비구조단백질을 코딩하는 유전자를 대장균에서 발현시켜 제조한 재조합 3ABC 비구조단백질 항원 및 3ABC 비구조단백질에 특이적인 단클론항체를 제조하고 이를 이용한 효소결합면역측정법을 실시함으로써 백신접종축과 야외감염축을 신속하고 정확하게 감별할 수 있는 구제역의 진단방법에 관한 것이다.