BACULOVIRUS EXPRESSION OF THE NUCLEOPROTEIN GENE OF MEASLES-VIRUS AND UTILITY OF THE RECOMBINANT PROTEIN IN DIAGNOSTIC ENZYME IMMUNOASSAYS

被引:141
作者
HUMMEL, KB [1 ]
ERDMAN, DD [1 ]
HEATH, J [1 ]
BELLINI, WJ [1 ]
机构
[1] CTR DIS CONTROL,NATL CTR INFECT DIS,DIV VIRAL & RICKETTSIAL DIS,ATLANTA,GA 30333
关键词
D O I
10.1128/JCM.30.11.2874-2880.1992
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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.
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页码:2874 / 2880
页数:7
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