Department of Immunology, Norman Bethune Medical College, Jilin University, Changchun 130021, China.
Vaccine. 2010 Apr 26;28(19):3435-9. doi: 10.1016/j.vaccine.2010.02.072. Epub 2010 Mar 1.
To distinguish the antibodies induced by Foot-and-mouth disease virus (FMDV) infection from those induced by vaccination, a recombinant N-terminal truncated FMDV non-structural protein (NSP) of 3AB, designated as r3aB, was constructed by deleting 80 amino acids displayed about 30% homology to transposase IS4 family protein of Escherichia coli, expressed in E. coli BL21 (DE3) and then purified. The r3aB was majorly expressed in soluble fraction and presented as homogeneous monomers after purification. Using r3aB as coating antigen, an indirect ELISA was established to specifically identify antibodies induced by FMDV infection but not those induced by vaccination. Compared with 3AB, r3aB was more specific to catch antibodies against NSP. The performance of this assay was validated by two commercial FMDV NSP ELISA kits. The result suggested that the r3aB coated ELISA could be developed into a kit to better distinguish between infected and vaccinated cattle.
为了区分口蹄疫病毒(FMDV)感染诱导的抗体与疫苗接种诱导的抗体,构建了一种截短的 3AB 区 N 端非结构蛋白(NSP)重组蛋白 r3aB,该蛋白缺失了与大肠杆菌转座酶 IS4 家族蛋白具有约 30%同源性的 80 个氨基酸,在大肠杆菌 BL21(DE3)中表达并纯化。r3aB 主要以可溶形式表达,经纯化后呈均一的单体。用 r3aB 作为包被抗原,建立了一种间接 ELISA 方法,该方法可特异性识别 FMDV 感染诱导的抗体,而不识别疫苗接种诱导的抗体。与 3AB 相比,r3aB 对 NSP 的抗体具有更高的特异性。该检测方法的性能已通过两种商业化的 FMDV NSP ELISA 试剂盒进行了验证。结果表明,r3aB 包被的 ELISA 可开发成试剂盒,以更好地区分感染牛和接种疫苗牛。