Development and evaluation of NS1 protein based i-ELISA for serodiagnosis of Bluetongue disease

Abstract

Bluetongue (BT) is an important viral disease of small ruminants and causes major economic losses in India. For serodiagnosis of BT, recombinant antigen-based assays are considered as alternatives to conventional approaches such as serum neutralization test. The present study aimed at the development of an indirect- enzyme-linked immunosorbent assay (i-ELISA) using bluetongue (BTV) recombinant NS1. NS1 gene of BTV encoding a mature NS1 protein was produced as a recombinant protein (rNS1) in Escherichia coli. The immunogenic potential of purified rNS1 was confirmed by detecting specific NS1 antibody responses in sera collected from immunized sheep using ELISA. An i-ELISA based on rNS1 was developed and optimized. In comparison to a commercially available c-ELISA kit, in the detection of BTV specific antibodies, this new assay exhibited a diagnostic sensitivity of 91.2%. This study highlighted the potential utility of rNS1 protein as a safe and novel diagnostic reagent in comparison to live virus antigen, in the development of serodiagnostic assay for surveillance of BTV infection in sheep and goats.