Optimisation of indirect elisa by comparison of different antigen preparations for detection of antibodies against schmallenberg [1] [2] virus

Abstract

Schmallenberg virus (SBV) infection, discovered in 2011, was reported in Europe including Turkey, Africa and recently in some Asian countries. Commercial enzyme-linked immunosorbent assay (ELISA) kits were widely used by researchers in many epidemiological studies and SBV diagnosis. The aim of this study was to optimise indirect in-house ELISA that is based on different antigen preparations of cell-culture derived whole SBV particle. Antigen preparations were maintained with various methods: PEG precipitation, ultracentrifugation, dialysis, and antigen inactivation. Following antigen optimisation, steps of antigen coating, blocking, conjugate and stop solution were optimised and in-house ELISA was compared to commercial indirect SBV ELISA kit. The best result in ELISA antigen preparation for SBV was gained by 30% PEG purification method followed by formaldehyde inactivation. Although results of this study demonstrated that in-house ELISA for detection of SBV specific antibodies was equally sensitive and specific as commercial kit, purified SBV antigen based in-house ELISA development could increase S/P ratios.