摘要
为建立EBOV抗体间接ELISA检测方法,以纯化后的EBOV糖蛋白作为包被抗原,HRP标记山羊抗马IgG为二抗,EBOV阳性马血清和阴性马血清分别为阳、阴性对照,优化反应条件,以EBOV病毒样颗粒免疫获得的高免马血清评价其特异性和敏感性,并将检测结果与基于假病毒的中和抗体检测结果相比较,进一步评价EBOV抗体ELISA检测方法在EBOV免疫血清抗体水平检测中的应用。优化后的间接ELISA方法可特异性检测EBOV抗体,与西尼罗病毒等的阳性血清均不发生反应,检测结果与基于假病毒的中和抗体检测结果相平行。批内、批间试验变异系数均小于8%。本研究成功建立了EBOV抗体间接ELISA检测方法,为EBOV免疫血清抗体水平检测提供了一种简便、快速的检测方法。
An indirect ELISA which can detect EBOV antibody was established using the purified the envelope glycoprotein as coating antigen.We use the HRP labeled antibody as the secondary antibody,the positive and negative horse sera as controls.After the reaction conditions were optimized,we analyzed the specificity,sensitivity of the indirect ELISA using serum of horse which immunized with EBOV virus like particles.For further evaluation of application of the ELISA,the results were compared to the neutralization assay based on pseudo-type EBOV.The indirect ELISA could detect the antibody against EBOV specifically and showed no cross-reaction with thepositive sera of West Nile Fever et al.The indirect ELISA also showed high coincidence with the neutralization assay based on pseudo-type EBOV in evaluation of the immune response after immunization.The coefficient of variation of inter-batch and intra-batch duplicative tests was less than 8%.The indirect ELISA was successfully developed.It provides an effective and convenient method to kinetic detection of antibody against EBOV after immunization.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2016年第3期615-619,共5页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家新药创制重大专项(2015ZX09102025)
