Lin Yafang, Zhou Lujia, Xiao Changguang, Li Zongjie, Liu Ke, Li Beibei, Shao Donghua, Qiu Yafeng, Ma Zhiyong, Wei Jianchao
Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China.
Front Microbiol. 2024 May 10;15:1359970. doi: 10.3389/fmicb.2024.1359970. eCollection 2024.
Porcine Reproductive and Respiratory Syndrome virus (PRRSV) causes high abortion rates in gestating sows and stillbirths, as well as high piglet mortality, seriously jeopardizing the pig industry in China and worldwide.
In this study, an infectious clone containing the full-length genome of NADC34-like PRRSV was constructed for the first time using reverse genetic techniques. The gene was amplified segmentally onto a plasmid, transfected into BHK-21 cells, and the transfected supernatant was harvested and transfected into PAM cells, which showed classical cytopathic effects (CPE).
The virus rJS-KS/2021 was successfully rescued which could be demonstrated by Western Blot and indirect immunofluorescence assays. Its growth curve was similar to the original strain. Replace the 5'UTR and 3'UTR of rJS-KS/2021 with 5'UTR and 3'UTR of HP-PRRSV (strain SH1) also failed to propagate on MARC-145.
In this study, an infectious clone of NADC34-like was constructed by reverse genetics, replacing the UTR and changing the cellular tropism of the virus. These findings provide a solid foundation for studying the recombination of different PRRSVs and the adaption of PRRSVs on MARC-145 in the future.
猪繁殖与呼吸综合征病毒（PRRSV）可导致妊娠母猪的高流产率和死胎，以及仔猪的高死亡率，严重危害中国乃至全球的养猪业。
在本研究中，首次利用反向遗传技术构建了包含NADC34样PRRSV全长基因组的感染性克隆。该基因被分段扩增到质粒上，转染至BHK-21细胞，收集转染后的上清液并转染至PAM细胞，PAM细胞呈现出典型的细胞病变效应（CPE）。
成功拯救出病毒rJS-KS/2021，这可通过蛋白质免疫印迹法和间接免疫荧光测定法得到证实。其生长曲线与原始毒株相似。用HP-PRRSV（毒株SH1）的5'UTR和3'UTR替换rJS-KS/2021的5'UTR和3'UTR后，该病毒也无法在MARC-145细胞上增殖。
在本研究中，通过反向遗传学构建了NADC34样的感染性克隆，替换UTR并改变了病毒的细胞嗜性。这些发现为今后研究不同PRRSV之间的重组以及PRRSV在MARC-145细胞上的适应性提供了坚实的基础。
