Czerny C P, Wagner K, Gessler K, Mayr A, Kaaden O R
Institute for Medical Microbiology, Infectious and Epidemic Diseases, Ludwig-Maximilians-University, Munich, Germany.
Vet Microbiol. 1996 Oct;52(3-4):185-200. doi: 10.1016/s0378-1135(96)00072-7.
A double sandwich blocking-ELISA using a genus-specific neutralizing monoclonal antibody (MAb) against the vaccinia virus 32 kD adsorption protein (D8L open reading frame: ORF) was developed to detect orthopoxvirus (OPV) antibodies in sera. A collection of 2173 feline serum samples was examined in an epidemiological study. The blocking-ELISA revealed 44 (2%) sera with positive titres of 1:2-1:256. ELISA results were confirmed by the plaque-reduction test. A close correlation between titres of both assays could be observed (r = 0.986). In general, the sensitivity of the blocking ELISA was two to four times higher. Neutralizing OPV-antibodies were found in nine sera with ELISA-titres > 1:4. Antibody specificity to OPV was also demonstrated by Western blotting analysis with selected feline sera. The epidemiographical distribution of the ELISA-positive sera and case histories of 37 seropositive cats available from the referring veterinarians are demonstrated. The blocking-ELISA enables a rapid serological diagnosis and can be used in veterinary and human medicine. It allows OPV-antibody screening in human and other animal species.
开发了一种双夹心阻断酶联免疫吸附测定法（ELISA），该方法使用针对痘苗病毒32 kD吸附蛋白（D8L开放阅读框：ORF）的属特异性中和单克隆抗体（MAb）来检测血清中的正痘病毒（OPV）抗体。在一项流行病学研究中检测了2173份猫血清样本。阻断ELISA检测出44份（2%）血清的阳性滴度为1:2至1:256。ELISA结果通过蚀斑减少试验得到证实。可以观察到两种检测方法的滴度之间有密切相关性（r = 0.986）。一般来说，阻断ELISA的灵敏度高两到四倍。在9份ELISA滴度>1:4的血清中发现了中和性OPV抗体。用选定的猫血清进行的蛋白质印迹分析也证明了抗体对OPV的特异性。展示了ELISA阳性血清的流行病学分布以及从转诊兽医处获得的37只血清阳性猫的病史。这种阻断ELISA能够实现快速血清学诊断，可用于兽医和人类医学。它允许在人类和其他动物物种中进行OPV抗体筛查。
