Wernike Kerstin, Brocchi Emiliana, Cordioli Paolo, Sénéchal Yann, Schelp Christian, Wegelt Anne, Aebischer Andrea, Roman-Sosa Gleyder, Reimann Ilona, Beer Martin
Institute of Diagnostic Virology, Friedrich-Loeffler-Institut (FLI), Suedufer 10, 17493, Greifswald - Insel Riems, Germany.
Istituto Zooprofilattico Sperimentale della Lombardia e dell'Emilia Romagna, Via Bianchi 7, 25125, Brescia, Italy.
Vet Res. 2015 Mar 11;46:27. doi: 10.1186/s13567-015-0165-4.
A panel of monoclonal antibodies (mAbs) specific for the nucleocapsid (N) protein or the glycoprotein Gc of Schmallenberg virus (SBV), a novel member of the Simbu serogroup (genus Orthobunyavirus, family Bunyaviridae), was produced and used to analyze antigenic differences among members of this serogroup. Reactivity with various SBV-isolates and other Simbu serogroup viruses was assessed by an indirect immunofluorescence test and by immunoblotting. The Gc-specific mAbs detected different SBV isolates as well as two closely related members of the Simbu serogroup. In addition, one mAb showed a highly specific reactivity with the homologous SBV strain only. Based on their differing reactivity with different SBV-strains, these antibodies represent a valuable novel tool to rapidly determine the phenotype of new SBV isolates. In contrast, the N-specific mAbs showed a broad reactivity spectrum and detected not only all the tested SBV-isolates, but also several other viruses of the Simbu serogroup. One out of these mAbs even recognized all of the tested Simbu serogroup viruses in the indirect immunofluorescence assay. In order to further characterize the N-specific antibodies, PepScan analysis was performed and a specific epitope could be identified. In summary, the newly generated mAbs showed differing pan-Simbu virus-, pan-SBV- as well as SBV-isolate-specific reactivity patterns. Thus, they represent valuable tools for the development of novel antigen and antibody detection systems either specific for SBV or, in a broader approach, for the pan-Simbu serogroup diagnostics.
制备了一组针对施马伦贝格病毒(SBV)核衣壳(N)蛋白或糖蛋白Gc的单克隆抗体(mAb),该病毒是辛布血清群(正布尼亚病毒属,布尼亚病毒科)的一个新成员,并用于分析该血清群成员之间的抗原差异。通过间接免疫荧光试验和免疫印迹法评估了与各种SBV分离株及其他辛布血清群病毒的反应性。Gc特异性mAb检测到不同的SBV分离株以及辛布血清群的两个密切相关成员。此外,一种mAb仅与同源SBV株表现出高度特异性反应。基于它们与不同SBV株的不同反应性,这些抗体是快速确定新SBV分离株表型的有价值的新工具。相比之下,N特异性mAb表现出广泛的反应谱,不仅检测到所有测试的SBV分离株,还检测到辛布血清群的其他几种病毒。其中一种mAb在间接免疫荧光试验中甚至识别了所有测试的辛布血清群病毒。为了进一步表征N特异性抗体,进行了肽扫描分析,并鉴定了一个特定表位。总之,新产生的mAb表现出不同的泛辛布病毒、泛SBV以及SBV分离株特异性反应模式。因此,它们是开发针对SBV或更广泛地针对泛辛布血清群诊断的新型抗原和抗体检测系统的有价值工具。