蓝舌病病毒4型特异性竞争ELISA检测方法的建立

Establishment of two competitive ELISAs for specific detection of bluetongue virus serotype 4

摘要:

旨在建立蓝舌病病毒4型 (BTV-4) 特异性抗体ELISA检测方法，为蓝舌病的免疫学诊断提供新的技术。利用制备的两株抗4型BTV VP2蛋白的单克隆抗体4A-1G7和4B-1B6，建立BTV-4特异性竞争ELISA抗体检测方法。利用该方法同时对50份羊和牛BTV阴性血清进行检测，分别确定两种方法阻断率临界值为49%和40%。利用标准阳性血清检测的试验结果表明，该方法的敏感性、特异性和重复性符合OIE通用标准。同时，4A-1G7和4B-1B6两种竞争ELISA方法联合作用，可以检测感染4、18和20型BTV的血清。研究结果为建立以上各型BTV的检测方法提供了基础。

Abstract:

To develop a clinical diagnosis technique for bluetongue virus infection, we established serotype-specific methods to detect serotype 4 of bluetongue virus (BTV-4). Two monoclonal antibodies (mAbs) against VP2 protein of BTV-4, named 4A-1G7 and 4B-1B6, were used as competitive antibodies in the competitive enzyme-linked immunosorbent assays (C-ELISA). We detected 50 negative serum samples from sheep, goats and cattle by C-ELISA. The cut-off values of 4A-1G7 and 4B-1B6 mAbs were 49% and 40%, respectively. The results of the sensitivity, specificity and repeatability by detecting standard positive serum, were consistent with the general standard of Office International Des Epizooties. Furthermore, serum samples of BTV-4, BTV-18 and BTV-20 infection could be screened out through the combined C-ELISAs by 4A-1G7 and 4B-1B6 mAbs. Thus, this technique may diagnose BTV-4, BTV-18 and BTV-20 infections.