Isolation and Characterization of Encephalomyocarditis Virus from Dogs in China

Ya-Kun Luo,Qing-Hai Tang,Li-Jun Shi,Ling Zhou,Lin Liang,Shang-Jin Cui,Ying-Ying Cong,Wen-Cheng Lin

doi:10.1038/s41598-017-00435-x

Abstract

Encephalomyocarditis virus (EMCV) is as a potential zoonotic agent with a wide host range. Here, we describe an EMC virus isolate, identified as EMCV C15, which was successfully obtained from the serum of dogs from animal hospitals. Virus production in cell culture was confirmed by EMCV-specific real-time RT-PCR, indirect immunofluorescence assays and electron microscopy. In addition, the open reading frame sequence (ORF) of the EMCV C15 virus was determined. From sequence comparison and phylogenetic analysis among 24 reference EMCV strains, it appears that the EMCV C15 strain is closely genetically related to strain BEL2887A/91 (>99.0% nucleotide identity). In artificially challenged dogs, the heart and brain were important targets of EMCV C15. This study provides genetic and pathogenic characterization of the EMCV C15 strain isolated in Beijing and calls for sustained surveillance of EMCV infection in China to support better prevention and control of the disease.

Highlights

Encephalomyocarditis virus (EMCV) is a small non-enveloped virus with a positive single-stranded genome, approximately 7.8 kb in length with a large open reading frame (ORF)
Western blot analysis demonstrated that capsid protein VP1 expression was similar in BHK-21 cells infected with EMCV-C15 or EMCV HB10, which was used as a positive control (Fig. 3)
These results show that EMCV-C15 and EMCV HB10 are indistinguishable with regard to viral replication and spreading in BHK-21 cells and that the monoclonal antibody recognizes an epitope common to both strains

Summary

Introduction

EMCV is a small non-enveloped virus with a positive single-stranded genome, approximately 7.8 kb in length with a large open reading frame (ORF). The ORF codes for a polyprotein that comprises both non-structural and structural elements divided into three primary precursor molecules—P1, P2 and P3—encoding for 11 distinct proteins[15]. The function of the EMCV proteins has often been assigned by virtue of their similarity to their well-studied counterparts poliovirus (PV), Theiler’s murine encephalomyelitis virus (TMEV) and foot and mouth disease virus (FMDV). This study describes the isolation, identification and characterization of an EMC virus, EMCV C15, from dogs. The ORF sequences of the EMCV C15. Isolate were compared with the ORF of 24 EMCV strains. In dogs artificially challenged with EMCV C15, the heart and brain were important targets for the virus and exhibited a viral load of over 105 gene copies. This study characterizes the molecular evolution of EMCV C15 in China and provides a reference for future studies on EMCV control and prevention

Methods

Results

Conclusion