Design and assessment of a double antigen indirect ELISA for lumpy skin disease surveillance in India

Lumpy skin disease (LSD), caused by the lumpy skin disease virus of the genus Capripoxvirus, is rapidly emerging across most countries in Asia. Recently, LSD has been linked to very high morbidity and mortality rates. Until 2019, India remained free of LSD, resulting in a lack of locally developed diagnostic kits, biologicals, and other tools necessary for managing the disease in a country with such a large livestock population. Therefore, this study aimed to design and validate an indigenous and cost-effective in-house ELISA for large-scale screening of cattle samples for antibodies to LSDV. The viral major open reading frames ORF 095 and ORF 103 encoding virion core proteins were expressed in a prokaryotic system and the recombinant antigen cocktail was used for optimization and validation of an indirect ELISA (iELISA). The calculated relative diagnostic sensitivity and diagnostic specificity of the iELISA were 96.6 % and 95.1 %, respectively at the cut-off percent positivity (PP≥50 %). The in-house designed double-antigen iELISA was found effective to investigate the seroprevalence of LSDV in various geographical regions of India. •An indigenous and cost-effective in-house ELISA for the large-scale screening of cattle samples for the antibodies to LSDV was designed and validated.•Viral ORF 095 and ORF 103 core proteins were expressed in a prokaryotic system; and the recombinant antigen cocktail optimized an iELISA.•The iELISA showed 96.6 % sensitivity and 95.1 %, specificity at a cut-off percent positivity (PP≥50 %).•The in-house double-antigen iELISA effectively investigated the seroprevalence of LSDV across various regions of India.