Characterization of a novel bat-HKU2-like swine enteric alphacoronavirus (SeACoV) infection in cultured cells and development of a SeACoV infectious clone

Yong-Le Yang; Qi-Zhang Liang; Shu-Ya Xu; Evgeniia Mazing; Guo-Han Xu; Lei Peng; Pan Qin; Bin Wang; Yao-Wei Huang

doi:10.1016/j.virol.2019.08.006

Characterization of a novel bat-HKU2-like swine enteric alphacoronavirus (SeACoV) infection in cultured cells and development of a SeACoV infectious clone

Virology. 2019 Oct:536:110-118. doi: 10.1016/j.virol.2019.08.006. Epub 2019 Aug 9.

Authors

Yong-Le Yang¹, Qi-Zhang Liang¹, Shu-Ya Xu¹, Evgeniia Mazing¹, Guo-Han Xu¹, Lei Peng¹, Pan Qin¹, Bin Wang¹, Yao-Wei Huang²

Affiliations

¹ Institute of Preventive Veterinary Medicine and Key Laboratory of Animal Virology of Ministry of Agriculture, Department of Veterinary Medicine, Zhejiang University, Hangzhou, 310058, Zhejiang, China.
² Institute of Preventive Veterinary Medicine and Key Laboratory of Animal Virology of Ministry of Agriculture, Department of Veterinary Medicine, Zhejiang University, Hangzhou, 310058, Zhejiang, China. Electronic address: yhuang@zju.edu.cn.

Abstract

Swine enteric alphacoronavirus (SeACoV), also known as swine acute diarrhea syndrome coronavirus (SADS-CoV), belongs to the species Rhinolophus bat coronavirus HKU2. Herein, we report on the primary characterization of SeACoV in vitro. Four antibodies against the SeACoV spike, membrane, nucleocapsid and nonstructural protein 3 capable of reacting with viral antigens in SeACoV-infected Vero cells were generated. We established a DNA-launched SeACoV infectious clone based on the cell adapted passage-10 virus and rescued the recombinant virus with a unique genetic marker in cultured cells. Six subgenomic mRNAs containing the leader-body junction sites, including a bicistronic mRNA encoding the accessory NS7a and NS7b genes, were experimentally identified in SeACoV-infected cells. Cellular ultrastructural changes induced by SeACoV infection were visualized by electron microscopy. The availability of the SeACoV infectious clone and a panel of antibodies against different viral proteins will facilitate further studies on understanding the molecular mechanisms of SeACoV replication and pathogenesis.

Keywords: Electron microscopy (EM); Infectious clone; Subgenomic mRNAs; Swine enteric alphacoronavirus (SeACoV); Viral antibodies.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alphacoronavirus / genetics*
Alphacoronavirus / metabolism
Alphacoronavirus / pathogenicity
Animals
Antibodies, Viral / biosynthesis
Antibodies, Viral / chemistry*
Antigens, Viral / chemistry*
Antigens, Viral / immunology
Base Sequence
Cell Membrane / ultrastructure
Cell Membrane / virology
Chiroptera
Chlorocebus aethiops
Clone Cells
Coronavirus Infections / diagnosis
Coronavirus Infections / veterinary*
Coronavirus Infections / virology
DNA, Complementary / genetics
DNA, Complementary / metabolism
Microscopy, Electron
Nucleocapsid / chemistry
Nucleocapsid / immunology
RNA, Messenger / genetics*
RNA, Messenger / metabolism
RNA, Viral / genetics*
RNA, Viral / metabolism
RNA-Dependent RNA Polymerase / chemistry
RNA-Dependent RNA Polymerase / immunology
Rabbits
Spike Glycoprotein, Coronavirus / chemistry
Spike Glycoprotein, Coronavirus / immunology
Swine
Swine Diseases / diagnosis
Swine Diseases / virology
Vero Cells
Viral Nonstructural Proteins / chemistry
Viral Nonstructural Proteins / immunology
Virus Replication

Substances

Antibodies, Viral
Antigens, Viral
DNA, Complementary
RNA, Messenger
RNA, Viral
Spike Glycoprotein, Coronavirus
Viral Nonstructural Proteins
RNA-Dependent RNA Polymerase

Supplementary concepts

Swine acute diarrhea syndrome coronavirus