A novel competitive ELISA for detection of antibodies against bovine viral diarrhea virus infection.

Diagnosis of bovine viral diarrhea (BVD) relies on the detection of antibodies against its viral causing agent, bovine viral diarrhea virus (BVDV). Here, we designed a novel competitive ELISA (cELISA) using the most immunogenic part of BVDV nonstructural protein 3 (NS), as a single ELISA recombinant antigen, along with a monoclonal antibody to detect antibodies against BVDV in sera of infected animals. Hence, 197 serum samples were tested by this cELISA and the results were compared to the results obtained from virus neutralization test (VNT) as the gold standard method for diagnosis of BVD.

Simple Indirect Enzyme-Linked Immunosorbent Assay to Detect Antibodies Against Bovine Viral Diarrhea Virus, Based on Prokaryotically Expressed Recombinant MBP-NS3 Protein.

Background: Bovine viral diarrhea (BVD) is an economically important disease of cattle distributed worldwide. Diagnosis of BVD relies on laboratory-based detection of its viral causing agent or virus specific antibodies and the most common laboratory method for this purpose is Enzyme-Linked Immunosorbent Assay (ELISA).

Objectives: The current study was aimed to develop a simple indirect ELISA to detect antibodies against Bovine Viral Diarrhea Virus (BVDV) in the sera of infected cattle.

Epitope mapping of bovine viral diarrhea virus nonstructural protein 3.

Six consecutive overlapped coding regions (F1-F6) of whole NS3 molecule of bovine viral diarrhea virus (BVDV) were cloned into pMAL-c2X plasmid vector and expressed in Escherichia coli cells (BL21 strain). The recombinant proteins were then purified by amylose resin to determine the most immunogenic domain(s) of the NS3 molecule. Evaluation of the recombinant proteins was carried out by indirect ELISAs using several bovine sera (previously characterized by virus neutralization test, a commercial ELISA kit, and a newly developed NS3-ELISA) and 6 monoclonal antibodies.