Czerny C P, Meyer H, Mahnel H
Zentralbl Veterinarmed B. 1989 Sep;36(7):537-46. doi: 10.1111/j.1439-0450.1989.tb00641.x.
Various combinations of polyclonal and neutralizing monoclonal antibodies (MAbs) of distinct specificity were evaluated as capture or detecting antibodies in an orthopox virus antigen ELISA. Acceptable results were achieved in an assay based on polyclonal antibodies. A 10 times higher sensitivity, however, was obtained using a combination of one monoclonal catching antibody reactive with viral envelope epitopes and polyclonal detection antibodies. This configuration proved to be superior in sensitivity to all others. Specificity was confirmed with 5 vaccinia virus strains (including one recombinant virus) and 8 species of orthopox viruses. Monkeypox and mousepox virus reacted exclusively in the polyclonal assay due to a lack of the specific epitope for the monoclonal antibody. The detection limit compared to the infectivity titer amounted to 10(3)-10(4) in the monoclonal/polyclonal and to 10(4)-10(5) TCID50/0.1 ml in the polyclonal combination. The correlation between infectivity- and ELISA-titer was demonstrated by a study of the replication cycle of the rabbitpox virus Uetrecht in the permanent rabbit kidney cell line RK-13. With the established ELISAs vaccinia virus could also be detected in organ suspensions of lethally infected NMRI-mice, depending on the level of infectivity.
在正痘病毒抗原酶联免疫吸附测定（ELISA）中，对具有不同特异性的多克隆抗体和中和单克隆抗体（MAb）的各种组合进行了评估，以作为捕获或检测抗体。基于多克隆抗体的检测取得了可接受的结果。然而，使用一种与病毒包膜表位反应的单克隆捕获抗体和多克隆检测抗体的组合，灵敏度提高了10倍。这种组合在灵敏度上被证明优于所有其他组合。用5种痘苗病毒株（包括1种重组病毒）和8种正痘病毒证实了其特异性。由于缺乏单克隆抗体的特异性表位，猴痘病毒和鼠痘病毒仅在多克隆检测中发生反应。与感染性滴度相比，单克隆/多克隆组合的检测限为10(3)-10(4)，多克隆组合的检测限为10(4)-10(5) TCID50/0.1 ml。通过对兔痘病毒Uetrecht在兔永久性肾细胞系RK-13中的复制周期研究，证明了感染性滴度与ELISA滴度之间的相关性。利用已建立的ELISA方法，根据感染水平，还可在致死感染的NMRI小鼠的器官悬液中检测到痘苗病毒。
