OPTIMIZATION OF THE MAIN PARAMETERS OF A LATERAL FLOW ASSAY BASED ON THE RECOMBINANT Р32 ANTIGEN OF LUMPY SKIN DISEASE VIRUS

Abstract

Lumpy skin disease is a highly contagious infectious transboundary disease of cattle, resulting in significant economic losses. The wide and rapid spread of lumpy skin disease to new territories emphasizes the objective need to improve diagnostic methods. The article outlines the findings of lateral flow assay parameter optimization for lumpy skin disease detection using recombinant P32 antigen. It was found that the optimal amount of protein for immobilization on 20 nm colloidal gold particles was 20 µg/mL. Bovine serum albumin at a final concentration of 0.25% was used to stabilize the conjugate. The optimal dilution of the conjugate was 1:2, and the concentration of recombinant protein and anti-species antibodies for application to the membrane was 500 μg/ml and 250 μg/ml, respectively. The colloidal gold conjugate interacted with antibodies and specifically reacted with the recombinant protein. No cross-reaction with other proteins was noticed at the same time. The created express test allowed for the detection of antibodies from immunized animals when the serum was diluted up to 1:200, however, 1:50 was the ideal ratio. The conjugate remained active after 90 days of storage at room temperature. The findings demonstrate the express method's potential for use in serological research for the prevention and control of the spread of lumpy skin disease.