检测马立克氏病毒抗体的酶联免疫吸附试验(ELISA)
AN ENZYME-LINKED IMMUNOSORBENT ASSAY FOR THE DETECTION OF ANTIBODIES TO MAREK’S DISEASE VIRUS
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摘要: 本文报导了检测马立克氏病毒(MDV)抗体的酶联免疫吸附试验(ELISA)。试验抗原为感染MDV的鸡胚成纤维(CEF)细胞,当阳性血清以1:400稀释时,最佳反应的细胞数为每孔5×104。我们比较了提纯的MDV抗原(PVAg)和感染MDV的全细胞抗原(WCAg),结果WcAg特异性反应强,非特异性反应弱。经WcAg包被的微量ELISA滴定板孔保存于4℃和-20℃,至少3个月不会降低抗原滴度。判定阳性血清的O.D.值为0.2。ELISA的敏感性比免疫荧光(IF)试验高20～40倍。抗血清对同型病毒抗原的滴度比对异型病毒抗原滴度高。这种ELISA方法适合于检测鸡血清中的MDV抗体和大量筛选马立克氏病毒单克隆抗体(MDV-McAb)。Abstract: A reproducible enzyme-linked immunosorbent assay(ELISA)using Marek’s disease virus(MDV)-infected cells for the detection of antibodies to MDV is described.The optimum number of MDV-infected chicken embryo fibroblasts(CEF)was 5 × 104/well,and test sera were positive at 1:400 dilutions.Compared with a purified virus preparation, MDV-infected CEF produced high specific and low nonspecific reactivities. wells coated with whole cells could be stored at 4 ℃ or-20℃ for at least 3 months without loss of reactivity, with antibody-negative sera, the cutoff absorbancy was 0.20 units.The ELISA was 20-to-40-fold more sensitive than indirect immunofluorescence.Homologous combinations of antisera in wells coated with CEF infected with different MDV serotypes were more reactive at higher dilutions than that were heterologous combinal ions. The procedure described is specific and suitable for large-scale screening of both chicken and monoclonal antibodies against MDV.
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