BACULOVIRUS EXPRESSION OF THE NUCLEOPROTEIN GENE OF MEASLES-VIRUS AND UTILITY OF THE RECOMBINANT PROTEIN IN DIAGNOSTIC ENZYME IMMUNOASSAYS
BACULOVIRUS EXPRESSION OF THE NUCLEOPROTEIN GENE OF MEASLES-VIRUS AND UTILITY OF THE RECOMBINANT PROTEIN IN DIAGNOSTIC ENZYME IMMUNOASSAYS
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DOI：
10.1128/jcm.30.11.2874-2880.1992
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发表时间：
1992-11-01
影响因子：
9.4
通讯作者：
BELLINI, WJ
中科院分区：
文献类型：
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作者：
HUMMEL, KB;ERDMAN, DD;BELLINI, WJ
产生了在多角体蛋白启动子的转录控制下表达麻疹病毒（埃德蒙斯顿疫苗株）核蛋白基因的重组杆状病毒。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示，表达的蛋白（B-MVN）与真实的病毒核蛋白共迁移，并被磷酸化。B-MVN蛋白被证明是与单克隆抗体在放射免疫沉淀反应，它是免疫原性的，在小鼠抗血清中引起认识到天然核蛋白。此外，评价了B-MVN蛋白作为酶免疫测定（EIA）中全病毒抗原的替代来源，用于检测麻疹病毒特异性免疫球蛋白M（IgM）和IgG抗体。以B-MVN蛋白为抗原的捕获IgM EIA在25例临床麻疹病例的18例（72%）急性期和所有恢复期标本中检测到特异性IgM抗体，120例对照标本的特异性超过99%。用B-MVN蛋白的间接IgG EIA在131份麻疹病毒抗体血清标本中的129份（98%）中检测到特异性IgG抗体，并且从测试268份额外血清标本中获得的结果与麻疹病毒中和抗体的相关性优于用商业EIA获得的结果。
A recombinant baculovirus that expresses the nucleoprotein gene of measles virus (Edmonston vaccine strain) under the transcriptional control of the polyhedrin promoter was generated. The expressed protein (B-MVN) comigrated with the authentic viral nucleoprotein as observed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and it was phosphorylated. The B-MVN protein proved to be reactive with monoclonal antibodies in radioimmunoprecipitations, and it was immunogenic, eliciting in mice antisera that recognized the native nucleoprotein. In addition, the B-MVN protein was evaluated as a replacement source of antigen for whole virus in enzyme immunoassays (EIAs) for detection of measles virus-specific immunoglobulin M (IgM) and IgG antibodies. A capture IgM EIA with the B-MVN protein as antigen detected specific IgM antibodies in 18 (72%) acute- and all convalescent-phase specimens from 25 clinical measles cases and exceeded 99% specificity with 120 control specimens. An indirect IgG EIA with the B-MVN protein detected specific IgG antibodies in 129 of 131 (98%) serum specimens with antibodies to measles virus, and results obtained from testing 268 additional serum specimens were better correlated with measles virus-neutralizing antibodies than those obtained with a commercial EIA.