Abstract
Porcine reproductive and respiratory syndrome (PRRS) is a highly contagious infectious disease caused by porcine reproductive and respiratory syndrome virus (PRRSV), which inflicts major economic losses on the global pig farming industry. Based on its similarity to highly pathogenic strains, the GDzj strain isolated in this study was predicted to be highly pathogenic. We therefore analyzed the pathogenicity of this strain experimentally in piglets. All piglets challenged with this virus experienced fever or high fever, loss of appetite, decreased food intake, daily weight loss, shortness of breath, and listlessness, and the necropsy results showed that they had experienced severe interstitial pneumonia. We then used the BAC system to construct a full-length cDNA infectious clone of GDzj, and the rescued virus displayed in vitro proliferation characteristics similar to those of the parental PRRSV strain. In summary, we successfully isolated a highly pathogenic PRRSV strain and constructed a full-length infectious cDNA clone from it, thereby providing an effective reverse genetics platform for further study of viral pathogenesis.
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Genetic data presented in this paper are publicly available via GenBank.
Abbreviations
- HP-PRRSV:
-
Highly pathogenic porcine reproductive and respiratory syndrome virus
- PRRS:
-
Porcine reproductive and respiratory syndrome
- ORFs:
-
Amino acid, aa; open reading frame
- NSP:
-
Non-structural protein
- BAC:
-
Bacterial artificial chromosome
- RT-PCR:
-
Reverse transcription polymerase chain reaction
- MOI:
-
Multiplicity of infection
- MCS:
-
Multiple cloning site
- FBS:
-
Fetal bovine serum
- IFA:
-
Immunofluorescence assay
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Acknowledgements
This work was supported by the National Key Research and Development Program of China (2018YFD0501100), Major animal disease prevention and control technology research and promotion projects (201817SY0002), Guangdong Province Key Field R&D Program Project (2019B020211003), The 2020 Youth Backbone Training Plan of Henan University of Higher Education (GaoJiao[2020]354), Key and Cultivation Discipline of Xinyang Agriculture and Forestry University (ZDXK201702), We thank Sandra Cheesman, PhD, from Liwen Bianji, Edanz Editing China (www.liwenbianji.cn/ac), for editing the English text of a draft of this manuscript.
Funding
This work was supported by the National Key Research and Development Program of China (2018YFD0501100), Major animal disease prevention and control technology research and promotion projects (201817SY0002), Guangdong Province Key Field R&D Program Project (2019B020211003), The 2020 Youth Backbone Training Plan of Henan University of Higher Education (GaoJiao[2020]354), Key and Cultivation Discipline of Xinyang Agriculture and Forestry University (ZDXK201702)
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SW, JD, and LY contributed equally to the work; SW, LY, and PZ performed the assays and performed the data analyses; TL, LZ, PL, and LW performed the sequence alignment; and JD, ZX and CS wrote the manuscript. All authors reviewed and approved the final form of the manuscript.
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All of the samples were collected according to the animal ethics regulations of the National Engineering Center for Swine Breeding Industry (NECSBI 2015-16).
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Wang, S., Liu, Y., Yu, L. et al. A strain of highly pathogenic porcine reproductive and respiratory syndrome virus: genomic characterization, pathogenicity, and construction of an infectious full-length cDNA clone. Arch Virol 166, 3127–3141 (2021). https://doi.org/10.1007/s00705-021-05212-w
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DOI: https://doi.org/10.1007/s00705-021-05212-w