Li Jitong, Zhu Tong, Lv Junfeng, Gao Yuehua, Hu Feng, Yu Kexiang, Song Minxun, Wang Jianlin, Wei Bai, Kang Min, Li Yufeng
Institute of Poultry Science, Shandong Provincial Key Laboratory of Immunity and Diagnosis of Poultry Diseases, Shandong Academy of Agricultural Sciences, Jinan, 250100, China.
College of Veterinary Medicine, Qingdao Agricultural University, Qingdao, 266109, China.
Sci Rep. 2025 Sep 26;15(1):33139. doi: 10.1038/s41598-025-17795-4.
A picornavirus strain of Duck/FC22/China/2017 (FC22) with an unclear taxonomic status was isolated in our laboratory in 2017. To study the biological properties of the virus, an infectious clone of the FC22 strain was successfully rescued by the infectious subgenomic amplicon method. The FC22 gene was amplified in two segments and fused with CMV and HDR after the addition of marker sites. The fusion fragment was transfected into LMH cells and passaged, and the obtained virus was named rFC22 and identified by PCR, IFA and electron microscopy. The biological characteristics of strain rFC22 were studied by inoculation with LMH cells, and the results showed that the TCID of rFC22 was essentially the same as that of its parent strain FC22 (1 × 10 TCID/mL) and that the growth curve of rFC22 was consistent with that of its parent strain FC22. The results indicated no significant difference in the biological characteristics between the rescued strain rFC22 and its parent strain FC22. In summary, our findings suggest that a duck-derived picornavirus was successfully rescued by ISA and the method was more convenient and more successful than alternative approaches.
2017年，我们实验室分离出一株分类地位不明的鸭源微小核糖核酸病毒毒株鸭/FC22/中国/2017（FC22）。为研究该病毒的生物学特性，采用感染性亚基因组扩增子法成功拯救出FC22毒株的感染性克隆。FC22基因分两段进行扩增，并在添加标记位点后与巨细胞病毒（CMV）和同源重组修复（HDR）融合。将融合片段转染至LMH细胞并传代，所得病毒命名为rFC22，并通过聚合酶链反应（PCR）、间接免疫荧光法（IFA）和电子显微镜进行鉴定。通过接种LMH细胞研究rFC22毒株的生物学特性，结果显示rFC22的半数组织培养感染剂量（TCID）与其亲本毒株FC22基本相同（1×10 TCID/mL），且rFC22的生长曲线与其亲本毒株FC22一致。结果表明，拯救毒株rFC22与其亲本毒株FC22的生物学特性无显著差异。综上所述，我们的研究结果表明，通过感染性亚基因组扩增子法成功拯救出一株鸭源微小核糖核酸病毒，且该方法比其他方法更简便、更成功。
