Rukhadze G G, Aliper T I, Sergeev V A
All-Union Research Institute of Experimental Veterinary Medicine VASKhNIL, Kuzminki, Moscow, USSR.
J Clin Microbiol. 1989 Aug;27(8):1754-8. doi: 10.1128/jcm.27.8.1754-1758.1989.
An enzyme-linked immunosorbent assay (ELISA) based on peplomer glycoprotein E2 was developed for the detection of antibodies to transmissible gastroenteritis virus (TGEV). Purified preparations of E2 were isolated by solubilization of the viral membrane with nonion detergent Nonidet P-40, followed by sucrose density gradient sedimentation. ELISA optical density values with E2 antigen significantly exceeded the indices when other TGEV protein or intact virion antigen was used. It was shown that a virus protein concentration in the E2 preparation of 500 ng per well is sufficient to sensitize the solid phase of microplates. A comparison of the ELISA and the virus neutralization test for the detection of TGEV antibodies was conducted. A significant correlation between the ELISA and the virus neutralization test was shown (r = 0.97). This serological test may be successfully used for various immunologic investigations.
开发了一种基于纤突糖蛋白E2的酶联免疫吸附测定(ELISA)法,用于检测猪传染性胃肠炎病毒(TGEV)抗体。通过用非离子去污剂Nonidet P-40溶解病毒膜,然后进行蔗糖密度梯度沉降,分离出纯化的E2制剂。与使用其他TGEV蛋白或完整病毒粒子抗原时相比,用E2抗原进行ELISA的光密度值显著更高。结果表明,每孔E2制剂中病毒蛋白浓度为500 ng足以使微孔板固相致敏。对ELISA法和病毒中和试验检测TGEV抗体进行了比较。结果显示ELISA法与病毒中和试验之间存在显著相关性(r = 0.97)。这种血清学检测方法可成功用于各种免疫学研究。