Bluetongue is an insect borne (Culicoides) viral disease of small ruminants. The virus blankets the globe with a wide serotypic variation, numbered from 1 to 28. In India 21 different serotypes have been reported to be circulating across the various agro-climatic zones of the country. Non-structural proteins (NSPs) of bluetongue virus have always remained ideal target for differentiation of infected from vaccinated animals. The current study is an extrapolation of our previous work where a novel fusion construct comprising of bluetongue viral segment NS1 and NS3 was successfully cloned, expressed, purified with an efficient strategy for its suitable implementation as a diagnostic antigen. In this study, the applicability of the fusion construct has been further evaluated and optimised for field applicability. The fusion construct used in an ELISA platform projected a relative diagnostic sensitivity and specificity of 98.1% and 95.5% respectively against a pre-established test panel. The rNS1-NS3 ELISA showed substantially good agreement with the commercial BTV antibody detection kit. Finally, the study brings together the diagnostic capability of two NSPs, which can be a handy tool for sero-surveillance of bluetongue.

蓝舌病是一种小反刍动物的虫媒 (Culicoides) 病毒性疾病。该病毒以广泛的血清型变异覆盖全球，编号从 1 到 28。在印度，据报道有 21 种不同的血清型在该国的各个农业气候带中传播。蓝舌病毒的非结构蛋白 (NSP) 一直是区分感染动物和接种疫苗的理想目标。目前的研究是我们之前工作的外推，其中包含蓝舌病毒片段 NS1 和 NS3 的新型融合构建体被成功克隆、表达、纯化，并采用有效策略将其作为诊断抗原合适地实施。在本研究中，融合构建体的适用性得到了进一步评估和优化，以实现现场适用性。ELISA 平台中使用的融合构建体相对于预先建立的测试组预测了 98.1% 和 95.5% 的相对诊断灵敏度和特异性。rNS1-NS3 ELISA 与商业 BTV 抗体检测试剂盒显示出非常好的一致性。最后，该研究汇集了两个 NSP 的诊断能力，这可以成为蓝舌病血清监测的便捷工具。