Key K F, DiCristina J, Gillespie J, Guenette D K, Meng X J
Center for Molecular Medicine and Infectious Diseases, Department of Biomedical Sciences and Pathobiology, College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061-0342, USA.
Arch Virol. 2007;152(7):1383-7. doi: 10.1007/s00705-007-0955-8. Epub 2007 Mar 15.
The recent construction of PRRSV infectious cDNA clones affords the opportunity for structural and functional studies of PRRSV genes. However, the inherent instability of the PRRSV genome, the requirement of cell culture propagation, and poor virus recovery have limited the usefulness of the PRRSV reverse genetics system for in vivo studies. Here, we report a unique strategy of infecting pigs by bypassing the traditional in vitro cell culture step required for in vivo studies. We demonstrate that inoculation of RNA transcripts of a PRRSV infectious cDNA clone directly into the lymph nodes and tonsils of pigs produces active PRRSV infection. The information from this study will have significant implications for the study of the molecular mechanism of PRRSV pathogenesis using the reverse genetics system.
猪繁殖与呼吸综合征病毒(PRRSV)感染性 cDNA 克隆的近期构建为 PRRSV 基因的结构和功能研究提供了机会。然而,PRRSV 基因组固有的不稳定性、细胞培养繁殖的要求以及病毒回收效率低,限制了 PRRSV 反向遗传系统在体内研究中的实用性。在此,我们报告了一种独特的策略,即绕过体内研究所需的传统体外细胞培养步骤来感染猪。我们证明,将 PRRSV 感染性 cDNA 克隆的 RNA 转录本直接接种到猪的淋巴结和扁桃体中可产生活跃的 PRRSV 感染。这项研究所得信息对于利用反向遗传系统研究 PRRSV 发病机制的分子机制具有重要意义。