College of Veterinary Medicine, Nanjing Agricultural University, Nanjing, 210095, People's Republic of China.
Vet Res. 2012 Jun 2;43(1):48. doi: 10.1186/1297-9716-43-48.
Peste des petits ruminants virus (PPRV) causes high mortality in goats and sheep and the disease has shown a greatly increased geographic distribution over the last 15 years. It is responsible for serious socioeconomic problems in some of the poorest developing countries. The ability to create recombinant PPRV would provide a useful tool for investigating the biology of the virus and the pathology of disease, as well as for developing new vaccines and diagnostic methods. Here we report the first successful rescue of recombinant PPRV from a full-length cDNA clone of the virus genome. Successful recovery of PPRV was achieved by using a RNA polymerase II promoter to drive transcription of the full-length virus antigenome. We have used this technique to construct a virus expressing a tracer protein (green fluorescent protein, GFP). The recombinant virus replicated as well as the parental virus and could stably express GFP during at least 10 passages. The newly established reverse genetics system for PPRV provides a novel method for constructing a vaccine using PPRV as a vector, and will also prove valuable for fundamental research on the biology of the virus. We found that our recombinant virus allowed more rapid and higher throughput assessment of PPRV neutralization antibody titer via the virus neutralization test (VNT) compared with the traditional method.
小反刍兽疫病毒（PPRV）可导致山羊和绵羊的高死亡率，并且在过去 15 年中，该疾病的地理分布范围大大扩大。它是一些最贫穷的发展中国家面临的严重社会经济问题的罪魁祸首。能够构建重组 PPRV 将为研究病毒的生物学和疾病病理学，以及开发新疫苗和诊断方法提供有用的工具。在这里，我们报告了首次从病毒基因组全长 cDNA 克隆中成功拯救重组 PPRV。通过使用 RNA 聚合酶 II 启动子驱动全长病毒抗原基因组的转录，成功地恢复了 PPRV。我们已经使用该技术构建了一种表达示踪蛋白（绿色荧光蛋白，GFP）的病毒。重组病毒的复制与亲本病毒一样，并且至少可以在 10 个传代中稳定表达 GFP。新建立的 PPRV 反向遗传学系统为使用 PPRV 作为载体构建疫苗提供了一种新方法，并且对于研究病毒的生物学也将非常有价值。我们发现，与传统方法相比，我们的重组病毒允许通过病毒中和试验（VNT）更快速且高通量地评估 PPRV 中和抗体滴度。
