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利用杆状病毒表达糖蛋白开发用于水泡性口炎病毒(VSV-IN)血清诊断的单克隆抗体连接酶联免疫吸附测定法。

Development of monoclonal antibody-linked ELISA for sero-diagnosis of vesicular stomatitis virus (VSV-IN) using baculovirus expressed glycoprotein.

作者信息

Kweon Chang Hee, Kwon Byung Joon, Kim In Joong, Lee Se Young, Ko Young Joon

机构信息

Animal Disease Diagnosis Division, National Veterinary Research and Quarantine Service, Ministry of Agriculture and Forestry, 480 Anyang, 6 Dong, Gyunggi Do, Republic of Korea.

出版信息

J Virol Methods. 2005 Dec;130(1-2):7-14. doi: 10.1016/j.jviromet.2005.05.023. Epub 2005 Aug 1.

Abstract

The gene encoding the envelope glycoprotein (GP) of vesicular stomatitis virus serotype, Indiana (VSV-IN), was expressed under the polyhedron promoter of baculovirus. The recombinant GP was applied as a diagnostic antigen for the detection of cattle and horse antibodies to VSV. In addition, the neutralizing monoclonal antibody (Mab) to GP of VSV-IN was used as trapping antibody in a Mab-linked indirect ELISA (MLI-ELISA) or detecting antibody in a Mab-linked competitive ELISA (MLC-ELISA). The diagnostic efficiencies of MLI-ELISA and MLC-ELISA were evaluated with currently available C-ELISA from OIE reference laboratory for vesicular stomatitis as a gold standard by using VSV-positive equine sera and negative bovine sera vaccinated against foot-and-mouth disease (FMD) in the field. When naturally infected equine sera and FMDV vaccinated bovine sera were tested, MLI-ELISA and MLC-ELISA showed relative sensitivities of 80% and 95% with relative specificity of 97% and 99%, respectively. However, both ELISAs cross-reacted with equine sera against New Jersey (VSV-NJ) serotype. The comparison of the two ELISAs revealed that MLC-ELISA was relatively more sensitive and specific than MLI-ELISA, indicating that MLC-ELISA can be applied to sero-diagnosis for VSV-IN infection.

摘要

水泡性口炎病毒印第安纳血清型(VSV-IN)包膜糖蛋白(GP)的编码基因在杆状病毒的多角体启动子下表达。重组GP被用作诊断抗原,用于检测牛和马针对VSV的抗体。此外,VSV-IN GP的中和单克隆抗体(Mab)在Mab连接的间接ELISA(MLI-ELISA)中用作捕获抗体,或在Mab连接的竞争ELISA(MLC-ELISA)中用作检测抗体。以OIE水泡性口炎参考实验室现有的C-ELISA作为金标准,使用现场VSV阳性马血清和接种口蹄疫(FMD)疫苗的阴性牛血清,评估MLI-ELISA和MLC-ELISA的诊断效率。当检测自然感染的马血清和接种FMDV的牛血清时,MLI-ELISA和MLC-ELISA的相对敏感性分别为80%和95%,相对特异性分别为97%和99%。然而,两种ELISA均与针对新泽西血清型(VSV-NJ)的马血清发生交叉反应。两种ELISA的比较表明,MLC-ELISA比MLI-ELISA相对更敏感和特异,表明MLC-ELISA可用于VSV-IN感染的血清学诊断。

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