以合成肽为抗原建立口蹄疫病毒非结构蛋白抗体检测ELISA试剂盒

Development of an ELISA for Detection of Foot-and-Mouth Nonstructural Protein Antibody with Synthetic Peptides

【目的】建立合成多肽抗原检测FMD NSP抗体的ELISA方法。【方法】固相法合成特异性FMDV NSP B细胞表位肽,将其偶联在BSA载体蛋白上,作为抗原包被在ELISA板上,制备检测抗FMDV NSP抗体的ELISA试剂盒,并对该试剂盒进行方法考核。【结果】抗原包被浓度选择2.5μg·mL-1;检测199份血清标本,与美国UBI商品试剂盒符合率达到96.48%,与国产3ABC ELISA比较,符合率为97.48%,显示极好的一致性。【结论】该合成肽ELISA试剂盒可以用以检测NSP抗体,从而鉴别FMD的自然感染与疫苗免疫,试剂盒特异性强,重复性好、稳定性高,操作简便。

[Objective] Developing an ELISA based on a synthetic peptide detecting foot-and-mouth disease virus（FMDV） nonstructural proteins （NSPs） antibody. [Method] A specifiC peptide according to FMDV NSPs B-cell epitopes was synthesized by a solid-phase method, and was conjugated with carder protein BSA. An ELISA for detecting FMDV NSPs antibody was developed by using the conjugating protein as the coating antigen. [Result] The optimal coating concentration of the antigen was determined as 2.5μg.mL^-1; The results of comparison of the assay with UBI NSP ELISA kit and national commercial 3ABC ELISA kit in detection of 199 serum samples showed that they were very coincident, and the agreement rate of them reached 96.48% and 97.48, respectively. [Conclusion] The developed ELISA using the synthetic peptide as coating antigen is specific, reproducible, stable and easy, and can be used to differentiate FMDV infected animals from immunized animals.