Mackay D K, Bulut A N, Rendle T, Davidson F, Ferris N P
Institute for Animal Health, Pirbright Laboratory, Ash Road, Pirbright, Woking, GU24 0NF, Surrey, UK.
J Virol Methods. 2001 Sep;97(1-2):33-48. doi: 10.1016/s0166-0934(01)00333-0.
A solid-phase competition ELISA has been developed to measure antibodies to foot-and-mouth disease (FMD) virus and has been validated using an extensive range of sera from cattle. The assay uses polyclonal antisera and inactivated purified 146S antigens of FMD virus and was compared with the liquid-phase blocking ELISA and the virus neutralisation test on a range of serum sets. When examining test sera at a 1:5 dilution with a cut-off point of 30% inhibition of reaction, the solid-phase competition ELISA was as sensitive as the liquid-phase blocking ELISA for sera from infected or vaccinated animals. The limit of detection of the solid-phase ELISA was similar to that of the liquid-phase assay and both tests had lower limit of detection (i.e. were able to detect lower amounts of antibody) than the virus neutralisation test. The specificity of the solid-phase ELISA was considerably higher than that of the liquid-phase blocking ELISA and almost equivalent to that of the virus neutralisation test. The assay thus retains the sensitivity of the liquid-phase blocking ELISA whilst being easier to use, more robust and specific, and therefore offers an improvement for FMD virus antibody detection.
已开发出一种固相竞争酶联免疫吸附测定法(ELISA)来检测口蹄疫(FMD)病毒抗体,并使用大量牛血清进行了验证。该测定法使用口蹄疫病毒的多克隆抗血清和灭活纯化的146S抗原,并与液相阻断ELISA以及一系列血清样本的病毒中和试验进行了比较。当以1:5稀释度检测测试血清且反应抑制临界点为30%时,固相竞争ELISA对于来自感染或接种动物的血清与液相阻断ELISA一样灵敏。固相ELISA的检测限与液相测定法相似,且两种检测方法的检测下限(即能够检测到的抗体量更低)均低于病毒中和试验。固相ELISA的特异性明显高于液相阻断ELISA,几乎与病毒中和试验相当。因此,该测定法保留了液相阻断ELISA的灵敏度,同时更易于使用、更稳定且更具特异性,从而为口蹄疫病毒抗体检测提供了改进。