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1.
Arch Virol ; 163(9): 2471-2476, 2018 Sep.
Article in English | MEDLINE | ID: mdl-29786119

ABSTRACT

Recombination occurs frequently between enteroviruses (EVs) which are classified within the same species of the Picornaviridae family. Here, using viral metagenomics, the genomes of two recombinant EV-Gs (strains EVG 01/NC_CHI/2014 and EVG 02/NC_CHI/2014) found in the feces of pigs from a swine farm in China are described. The two strains are characterized by distinct insertion of a papain-like protease gene from toroviruses classified within the Coronaviridae family. According to recent reports the site of the torovirus protease insertion was located at the 2C/3A junction region in EVG 02/NC_CHI/2014. For the other variant EVG 01/NC_CHI/2014, the inserted protease sequence replaced the entire viral capsid protein region up to the VP1/2A junction. These two EV-G strains were highly prevalent in the same pig farm with all animals shedding the full-length genome (EVG 02/NC_CHI/2014) while 65% also shed the capsid deletion mutant (EVG 01/NC_CHI/2014). A helper-defective virus relationship between the two co-circulating EV-G recombinants is hypothesized.


Subject(s)
Enterovirus Infections/veterinary , Enteroviruses, Porcine/genetics , Genome, Viral , Reassortant Viruses/genetics , Swine Diseases/epidemiology , Torovirus Infections/veterinary , Torovirus/genetics , Animals , Capsid Proteins/genetics , Capsid Proteins/metabolism , China/epidemiology , Endopeptidases/genetics , Endopeptidases/metabolism , Enterovirus Infections/epidemiology , Enterovirus Infections/virology , Enteroviruses, Porcine/classification , Enteroviruses, Porcine/metabolism , Farms , Feces/virology , Gene Deletion , Genetic Variation , Metagenomics/methods , Phylogeny , Prevalence , Reassortant Viruses/classification , Reassortant Viruses/metabolism , Recombination, Genetic , Swine , Swine Diseases/virology , Torovirus/classification , Torovirus/metabolism , Torovirus Infections/epidemiology , Torovirus Infections/virology , Viral Proteins/genetics , Viral Proteins/metabolism
2.
J Virol ; 90(8): 4067-4077, 2016 Apr.
Article in English | MEDLINE | ID: mdl-26865725

ABSTRACT

UNLABELLED: The receptor(s) for porcine sapelovirus (PSV), which causes diarrhea, pneumonia, polioencephalomyelitis, and reproductive disorders in pigs, remains largely unknown. Given the precedent for other picornaviruses which use terminal sialic acids (SAs) as receptors, we examined the role of SAs in PSV binding and infection. Using a variety of approaches, including treating cells with a carbohydrate-destroying chemical (NaIO4), mono- or oligosaccharides (N-acetylneuraminic acid, galactose, and 6'-sialyllactose), linkage-specific sialidases (neuraminidase and sialidase S), lectins (Maakia amurensislectin andSambucus nigralectin), proteases (trypsin and chymotrypsin), and glucosylceramide synthase inhibitors (dl-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol and phospholipase C), we demonstrated that PSV could recognize α2,3-linked SA on glycolipids as a receptor. On the other hand, PSVs had no binding affinity for synthetic histo-blood group antigens (HBGAs), suggesting that PSVs could not use HBGAs as receptors. Depletion of cell surface glycolipids followed by reconstitution studies indicated that GD1a ganglioside, but not other gangliosides, could restore PSV binding and infection, further confirming α2,3-linked SA on GD1a as a PSV receptor. Our results could provide significant information on the understanding of the life cycle of sapelovirus and other picornaviruses. For the broader community in the area of pathogens and pathogenesis, these findings and insights could contribute to the development of affordable, useful, and efficient drugs for anti-sapelovirus therapy. IMPORTANCE: The porcine sapelovirus (PSV) is known to cause enteritis, pneumonia, polioencephalomyelitis, and reproductive disorders in pigs. However, the receptor(s) that the PSV utilizes to enter host cells remains largely unknown. Using a variety of approaches, we showed that α2,3-linked terminal sialic acid (SA) on the cell surface GD1a ganglioside could be used for PSV binding and infection as a receptor. On the other hand, histo-blood group antigens also present in the cell surface carbohydrates could not be utilized as PSV receptors for binding and infection. These findings should contribute to the understanding of the sapelovirus life cycle and to the development of affordable, useful and efficient drugs for anti-sapelovirus therapy.


Subject(s)
Enteroviruses, Porcine/metabolism , Gangliosides/metabolism , N-Acetylneuraminic Acid/metabolism , Receptors, Virus/metabolism , Animals , Blood Group Antigens/metabolism , Carbohydrates/chemistry , Cell Line , HeLa Cells , Humans , N-Acetylneuraminic Acid/chemistry , Receptors, Virus/chemistry , Swine , Virus Attachment
3.
J Virol Methods ; 65(1): 111-21, 1997 Apr.
Article in English | MEDLINE | ID: mdl-9128868

ABSTRACT

Two polymerase chain reaction (PCR) assays are described for the detection of swine vesicular disease virus (SVDV) RNA, a reverse transcription PCR (RT-PCR) and a reverse transcription nested PCR (RT-nPCR). Both the RT-PCR and RT-nPCR were able to detect representative members of each of seven phylogenetically distinct groups of SVDV and gave negative results with a range of porcine enteroviruses and of viruses responsible for vesicular conditions in pigs. When combined with a commercial kit for rapid RNA extraction, the RT-PCR was useful for the detection of SVDV in samples of epithelium and faeces from animals with clinical SVD. The addition of a second amplification step to create a nested PCR (RT-nPCR) increased the sensitivity of the technique for the detection of viral RNA (vRNA) in SVDV infected tissue culture fluid by a factor of approximately 1,000, for 100 TCID50 for the RT-PCR to 0.1 TCID50 for RT-nPCR. When combined with a more elaborate extraction procedure for RNA, the RT-nPCR was considerably more sensitive than virus isolation in tissue culture for detecting SVDV in nasal swabs, tissues, and faeces collected from pigs between 7 days and 176 days after infection with a recent European isolate of SVDV. However, stringent conditions are necessary for carrying out the RT-nPCR to minimise the possibility of contamination.


Subject(s)
Enterovirus Infections/genetics , Enteroviruses, Porcine/genetics , Polymerase Chain Reaction/methods , RNA, Viral/metabolism , Swine Vesicular Disease/virology , Animals , Enterovirus Infections/veterinary , Enteroviruses, Porcine/metabolism , Polymerase Chain Reaction/veterinary , RNA, Viral/isolation & purification , RNA-Directed DNA Polymerase , Sensitivity and Specificity , Swine , Swine Vesicular Disease/diagnosis , Swine Vesicular Disease/genetics
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