A DIAGNOSTIC IMMUNOASSAY FOR NEWCASTLE-DISEASE VIRUS-BASED ON THE NUCLEOCAPSID PROTEIN EXPRESSED BY A RECOMBINANT BACULOVIRUS
A DIAGNOSTIC IMMUNOASSAY FOR NEWCASTLE-DISEASE VIRUS-BASED ON THE NUCLEOCAPSID PROTEIN EXPRESSED BY A RECOMBINANT BACULOVIRUS
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DOI：
10.1016/0166-0934(95)00074-7
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发表时间：
1995-11-01
影响因子：
3.1
通讯作者：
EMMERSON, P
中科院分区：
文献类型：
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作者：
ERRINGTON, W;STEWARD, M;EMMERSON, P
含有编码纽卡斯尔病病毒（阿尔斯特2C株）核衣壳（NP）蛋白的cDNA克隆的重组杆状病毒已被用于感染昆虫细胞（草地贪夜蛾）。观察到高水平的过表达NP蛋白，其占总细胞蛋白的高达40%，随后显示其具有抗原性。来源于感染的昆虫细胞的粗可溶性裂解物的核蛋白已被用于间接ELISA以检测鸡血清群中抗NDV抗体的存在。从这些试验中产生的数据表明，ELISA滴度和血凝抑制试验数据之间具有良好的相关性。该检测不受背景细胞蛋白的干扰，也不受与非NDV家禽病原体的交叉反应性的影响。此外，该测试没有产生假阳性读数。
A recombinant baculovirus containing a cDNA clone encoding the nucleocapsid (NP) protein of Newcastle disease virus (strain Ulster 2C) has been used to infect insect cells (Spodoptera frugiperda). High levels of overexpressed NP protein were observed, comprising up to 40% of total cellular protein, which were subsequently shown to be antigenic. Nucleoprotein derived from the crude soluble lysate of infected insect cells has been used in an indirect ELISA to detect the presence of anti-NDV antibodies in a cohort of chicken sera. Data produced from these tests indicated a good correlation between ELISA titre and haemagglutination inhibition test data. The test was not affected by interference from background cellular proteins nor by cross-reactivity with non-NDV poultry pathogens. Additionally, the test did not generate false-positive readings.