Isolation, molecular characterization, and phylogenetic analysis of porcine encephalomyocarditis virus strain HB10 in China

Lin, Wencheng; Liu, Yebing; Cui, Shangjin; Li, Hongmei

doi:10.1016/j.meegid.2012.04.008

Cited by 15 publications

(8 citation statements)

References 11 publications

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“…The complete 6,879-nt EMCV C15 ORF GenBank accession number (KU664327) encoding 2,292 amino acids with no insertions or deletions was examined. Multiple sequence alignment based on the ORF sequence of EMCV-C15 and other EMCV reference strains was performed using the Clustal X program 17 . The results showed the newly isolated virus (EMCV C15) shared the highest sequence similarity (99.0–99.75%) with GX0601, BJC3, NJ08, GS01, HB10 and BEL-2887A/91.…”

Section: Resultsmentioning

confidence: 99%

“…Then, serum samples were filtered through a 0.22-μm filter and used as an inoculum for EMC virus isolation. Isolation of EMCV C15 was attempted using BHK21 cells as previously described with some modifications 17 . Confluent BHK21 cells in 6-well plates were washed twice with postinoculation medium and inoculated with 300 μL of sample and 100 μL of postinoculation medium.…”

Section: Methodsmentioning

confidence: 99%

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Isolation and Characterization of Encephalomyocarditis Virus from Dogs in China

Luo

Liang

Tang

et al. 2017

Sci Rep

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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.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Isolation and Characterization of Encephalomyocarditis Virus from Dogs in China

Luo

Liang

Tang

et al. 2017

Sci Rep

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“…The pig is the mammal most likely to be impacted on with the disease [5, 6], but EMCV occurrence has also been reported in non-human primates and in a variety of domestic, captive and wild animals. Several lethal outbreaks have been described in zoos in Australia and the USA [2, 7, 8] and in Northern Italy [3].…”

Section: Introductionmentioning

confidence: 99%

Encephalomyocarditis virus infection in Macaca sylvanus and Hystrix cristata from an Italian rescue centre for wild and exotic animals

Cardeti

Mariano

Eleni

et al. 2016

Virol J

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BackgroundThe Encephalomyocarditis virus (EMCV) is a small, non enveloped, positive sense single-stranded RNA virus in the genus Cardiovirus, family Picornaviridae, with two known serotypes. It is spread worldwide and infects a huge range of vertebrate hosts with zoonotic potential for humans. The pig is the mammal most likely to be impacted on with the disease, but EMCV occurrence has also been reported in non-human primates and in a variety of domestic, captive and wild animals. Until now, human cases have been very rare and the risk appears to be almost negligible in spite of human susceptibility to the infection.Case presentationBetween September and November 2012 a fatal Encephalomyocarditis virus outbreak involving four Barbary macaques and 24 crested porcupines occurred at a rescue centre for wild and exotic animals in Central Italy. In this open-field zoo park located near Grosseto, Tuscany about 1000 animals belonging to different species, including various non-human primates were hosted at that time. Sudden deaths were generally observed without any evident symptoms or only with mild nonspecific clinical signs. The major gross change was characterised by grey-white necrotic foci in the myocardium and the same EMCV strain was isolated both in macaques and crested porcupines. Phylogenetic analysis has confirmed that only one EMCV strain is circulating in Italy, capable of infecting different animal species.ConclusionsThis report confirms the susceptibility of non-human primates to the EMCV infection and describes the disease in porcupine, a common wild Italian and African species. No human cases were observed, but given the zoonotic potential of EMCV these findings are of importance in the context of animal-human interface.

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“…However, no live attenuated vaccine is available. Recently, the EMCV-HB10 strain was isolated from the organs of dead piglets with acute myocarditis in northern China (Bai et al, 2012;Feng et al, 2015;Lin et al, 2012;Yuan et al, 2014). To date, the virulence and pathogenicity of this virus and the capacity of its genome to tolerate heterologous genes remain unclear.…”

Section: Discussionmentioning

confidence: 99%

An attenuated EMCV-HB10 strain acts as a live viral vector delivering a foreign gene

Huang

Zhang

et al. 2016

Journal of General Virology

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We successfully constructed a full-length cDNA infectious clone of the encephalomyocarditis virus (EMCV) HB10 strain and obtained a partially attenuated rEMCV-C 9 virus with a shorter poly(C) tract. Our results showed that the length of the EMCV-HB10 poly(C) tract was related to the pathogenicity of the EMCV-HB10 strain in vivo. Using pEMCV-C 9 as the backbone, we constructed the novel viral vector pC 9 -MCS-D2A by inserting a cDNA fragment containing a 127 amino acid deletion in the 2A protein, a primary cleavage cassette, a FLAG tag and a multiple cloning site (MCS) at the junction of VP1 and D2A. Additionally, the enhanced green fluorescent protein (egfp) gene was cloned into the MCS of pC 9 -MCS-D2A to test its capacity to express foreign proteins. Insertion of the egfp gene did not affect viral replication, and a decrease in EGFP expression was observed within five serial passages. Furthermore, we found that rC 9 -EGFP-D2A was avirulent in vivo, induced neutralizing antibody production and conferred protective immune responses against lethal challenge with EMCV in mice. Taken together, our results demonstrated that we had constructed an attenuated live vector based on an EMCV-HB10 strain with two modified critical virulence factors (the poly(C) tract and 2A protein) that could be used as a candidate live vaccine and a potential live viral vector for foreign antigen delivery.

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Isolation, molecular characterization, and phylogenetic analysis of porcine encephalomyocarditis virus strain HB10 in China

Cited by 15 publications

References 11 publications

Isolation and Characterization of Encephalomyocarditis Virus from Dogs in China

Isolation and Characterization of Encephalomyocarditis Virus from Dogs in China

Encephalomyocarditis virus infection in Macaca sylvanus and Hystrix cristata from an Italian rescue centre for wild and exotic animals

An attenuated EMCV-HB10 strain acts as a live viral vector delivering a foreign gene

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