Van Maanen C, Terpstra C
Central Veterinary Institute, Department of Virology, Lelystad, The Netherlands.
Biologicals. 1990 Oct;18(4):315-9. doi: 10.1016/1045-1056(90)90036-y.
A double antibody sandwich (DAS) enzyme-linked immunosorbent assay (ELISA) was developed to quantify 146S antigen of foot-and-mouth disease virus (FMDV) strain A10 Holland grown in suspension cultures of surviving bovine tongue epithelium. When virus harvests were incubated with trypsin--which affects VP1, the most immunogenic structural protein of FMDV--the concentration of 146S antigen as determined by ELISA was reduced by greater than 90%. Therefore, the test detected essentially only those virus particles with intact VP1. When the test was compared with the sucrose density gradient method, concentrations of 146S antigen correlated well (r = 0.87). The rate of variation in both tests was the same. In contrast to the sucrose density gradient method, the DAS-ELISA can simultaneously quantify 146S antigen in many samples, and also indicates when VP1 of 146S particles has disintegrated by the action of proteases.
开发了一种双抗体夹心（DAS）酶联免疫吸附测定（ELISA）法，用于定量在存活牛舌上皮悬浮培养物中生长的口蹄疫病毒（FMDV）A10荷兰株的146S抗原。当病毒收获物与胰蛋白酶孵育时（胰蛋白酶会影响FMDV免疫原性最强的结构蛋白VP1），ELISA测定的146S抗原浓度降低了90%以上。因此，该检测基本上只检测到那些VP1完整的病毒颗粒。当将该检测方法与蔗糖密度梯度法进行比较时，146S抗原浓度具有良好的相关性（r = 0.87）。两种检测方法的变化率相同。与蔗糖密度梯度法不同，DAS-ELISA可以同时定量许多样品中的146S抗原，还能表明146S颗粒的VP1何时因蛋白酶的作用而分解。
