Brown J M, Coates D M, Phillpotts R J
Chemical and Biological Defence Establishment, Salisbury, Wiltshire, UK.
J Virol Methods. 1996 Dec;62(2):143-51. doi: 10.1016/s0166-0934(96)02095-2.
Three monoclonal antibodies (Mabs) specific for the envelope (E) protein of flaviviruses were evaluated for use in an antigen capture ELISA. Three combinations of Mabs and a combination of polyclonal antibodies (Pabs) were evaluated in antigen capture ELISAs for their ability to detect 18 flaviviruses. The Mab ELISAs detected 50% of flavivirus antigens with a sensitivity between 1 and 9 x 10(4)/ng viral protein/ml, however, none of the ELISAs evaluated proved to be useful for generic detection of flaviviruses, being unable to detect tick-borne flaviviruses and some mosquito-borne flaviviruses. The inability of the ELISAs to detect tick-borne flaviviruses is thought to be due to the conformation of surface epitopes, which the Mabs were unable to recognise. This was again observed using recombinant TBE virus prM/E protein as antigen in direct and antigen capture ELISAs. The Mabs reacted with the prM/E protein when it was denatured by binding directly onto the solid phase, but the antibodies were unable to detect the native protein in antigen capture ELISAs. The antigen capture ELISAs evaluated in this study were considered to be unsuitable for the generic detection of flaviviruses, but may provide a sensitive diagnostic assay for specific flavivirus infection.
对三种针对黄病毒包膜（E）蛋白的单克隆抗体（Mab）进行了评估，以用于抗原捕获ELISA。在抗原捕获ELISA中评估了三种Mab组合以及一种多克隆抗体（Pab）组合检测18种黄病毒的能力。Mab ELISA检测到50%的黄病毒抗原，灵敏度在1至9×10⁴/ng病毒蛋白/毫升之间，然而，所评估的ELISA均未被证明对黄病毒的通用检测有用，无法检测蜱传黄病毒和一些蚊传黄病毒。ELISA无法检测蜱传黄病毒被认为是由于表面表位的构象，Mab无法识别。在直接ELISA和抗原捕获ELISA中使用重组蜱传脑炎病毒prM/E蛋白作为抗原时再次观察到这一点。当prM/E蛋白通过直接结合到固相上而变性时，Mab与prM/E蛋白发生反应，但在抗原捕获ELISA中抗体无法检测到天然蛋白。本研究中评估的抗原捕获ELISA被认为不适用于黄病毒的通用检测，但可能为特定黄病毒感染提供灵敏的诊断检测方法。
