East China Sea Fisheries Research Institute, Chinese Academy of Fisheries Science, 300 Jungong Road, Shanghai 200090, China.
Virus Res. 2010 Apr;149(1):51-5. doi: 10.1016/j.virusres.2009.12.012. Epub 2010 Jan 18.
Transmissible gastroenteritis virus (TGEV) is the causative agent of porcine transmissible gastroenteritis (TGE), characterized by high mortality and severely retarded growth in piglets that dramatically affects the porcine industry. Previously, we have identified two shRNA-expressing plasmids pEGFP-U6/P1 and pEGFP-U6/P2 that target RNA-dependent RNA polymerase (RdRP) gene of TGEV with more than 95% of virus inhibition in vitro. In this study, inhibition of the TGEV replication by pEGFP-U6/P1 and pEGFP-U6/P2 was tested in mini-pigs. SPF mini-pigs at 25 days old were injected with the shRNA-expressing plasmids and then infected with TGEV. The results from the analyses of clinical signs, histopathology, indirect immunofluorescence (IIF) and RT-PCR show that the two shRNA-expressing plasmids could significantly decrease the quantity of TGEV in different organs and protect mini-pigs from TGEV infection. These findings illustrate the prospect for TGEV-specific shRNAs to be new anti-TGEV agents.
传染性胃肠炎病毒(TGEV)是引起猪传染性胃肠炎(TGE)的病原体,其特征是仔猪死亡率高,生长严重迟缓,这对养猪业有重大影响。先前,我们已经鉴定了两种表达短发夹 RNA(shRNA)的质粒 pEGFP-U6/P1 和 pEGFP-U6/P2,它们针对 TGEV 的 RNA 依赖性 RNA 聚合酶(RdRP)基因,在体外抑制超过 95%的病毒。在这项研究中,我们在小型猪中测试了 pEGFP-U6/P1 和 pEGFP-U6/P2 对 TGEV 复制的抑制作用。25 天大的 SPF 小型猪被注射了表达 shRNA 的质粒,然后感染 TGEV。临床症状、组织病理学、间接免疫荧光(IIF)和 RT-PCR 分析的结果表明,两种表达 shRNA 的质粒可以显著降低不同器官中 TGEV 的数量,并保护小型猪免受 TGEV 感染。这些发现表明 TGEV 特异性 shRNA 有希望成为新型抗 TGEV 药物。