Nanobody-based competitive enzyme-linked immunosorbent assay for detecting antibodies of porcine reproductive and respiratory syndrome virus
文献类型： 外文期刊
作者： Sun, Mingxia 1 ; Cao, Dan 2 ; Zhao, Man 1 ; Zhu, Xulong 1 ; Sun, Yue 1 ; Wang, Qian 1 ; Zhang, Hongliang 1 ; Yang, Yongbo 1 ; Tian, Xiaoxiao 1 ; Wang, Haiwei 1 ; Cai, Xuehui 1 ; An, Tongqing 1 ;
作者机构： 1.Chinese Acad Agr Sci, Harbin Vet Res Inst, State Key Lab Vet Biotechnol, 678 Haping Rd, Harbin 150069, Peoples R China
2.Heilongjiang Acad Agr Sci, Soybean Res Inst, Harbin 150086, Peoples R China
3.Heilongjiang Prov Key Lab Vet Immunol, Harbin 150069, Peoples R China
关键词： Nanobody; PRRSV; Competitive ELISA; High specificity
期刊名称：INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES （ 影响因子：8.5； 五年影响因子：8.7 ）
ISSN： 0141-8130
年卷期： 2025 年 312 卷
页码：
收录情况： SCI
摘要： The pandemic of porcine reproductive and respiratory syndrome (PRRS) continues a global threat to the swine industry and causes huge economic losses. Large-scale serological surveillance, particularly through enzymelinked immunosorbent assay (ELISA), is an effective measure for PRRS control. Nanobodies have been used in diagnostic immunoassays due to their high affinity and specificity. In this study, two novel nanobodies against N protein of PRRSV were generated that exhibited ability to block the recognition of serum antibodies to N protein. A high-affinity nanobody was conjugated with horseradish peroxidase (HRP), then nanobody-based competitive ELISA (cELISA) was subsequently developed to detect antibodies against both genotypes 1 and 2 PRRSV. Notably, the developed cELISA exhibited enhanced sensitivity, specificity and reproducibility. The comparative analysis demonstrated a high agreement of 96.69 % between the cELISA and a commercial ELISA kit. Collectively, the developed nanobody-based cELISA can be used for epidemiological study of PRRSV infection and immune evaluations with advantages of simple operation and low costing.
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