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phylogenetic analysis of ns5b gene of classical swine fever virus isolates indicates plausible chinese origin of indian subgroup 2.2 viruses.twenty-three csfv isolates recovered from field outbreaks in various parts of india during 2006-2009 were used for genetic analysis in the ns5b region (409 nts). seventeen of these were studied earlier [16] in the 5'utr region. phylogenetic analysis indicated the continued dominance of subgroup 1.1 strains in the country. detailed analysis of a subgroup 2.2 virus indicated the plausible chinese origin of this subgroup in india and provided indirect evidence of routes of csfv movement within sout ...201121246270
processing and localization of the african swine fever virus cd2v transmembrane protein.the african swine fever virus (asfv)-encoded cd2v transmembrane protein is required for the hemadsorption of red blood cells around infected cells and is also required for the inhibition of bystander lymphocyte proliferation in response to mitogens. we studied the expression of cd2v by expressing the gene with a v5 tag downstream from the signal peptide near the n terminus and a hemagglutinin (ha) tag at the c terminus. in asfv-infected cells, a full-length glycosylated form of the cd2v protein, ...201121248037
human mxa protein inhibits the replication of classical swine fever virus.classical swine fever virus (csfv) has a spherical enveloped particle with a single stranded rna genome, the virus belonging to a pestivirus of the family flaviviridae is the causative agent of an acute contagious disease classical swine fever (csf). the interferon-induced mxa protein has been widely shown to inhibit the life cycle of certain rna viruses as members of the bunyaviridae family and others. interestingly, it has been reported that expression of mxa in infected cells was blocked by c ...201121255621
identification of a linear epitope on the capsid protein of classical swine fever virus.the capsid (c) protein of classical swine fever virus (csfv) is proposed to play an essential role in the replication and translation of the viral rna. in this study, a monoclonal antibody (mab) directed against the c protein was generated with the recombinant c protein expressed in escherichia coli as immunogen. ifa and ipma analysis showed that the native c protein of csfv virions was reactive to the mab. by truncating the c protein, we identified a linear epitope recognized by the mab, corres ...201121255622
interaction between core protein of classical swine fever virus with cellular iqgap1 protein appears essential for virulence in swine.here we show that iqgap1, a cellular protein that plays a pivotal role as a regulator of the cytoskeleton interacts with classical swine fever virus (csfv) core protein. sequence analyses identified residues within csfv core protein (designated as areas i, ii, iii and iv) that maintain homology to regions within the matrix protein of moloney murine leukemia virus (mmlv) that mediate binding to iqgap1 [embo j, 2006 25:2155]. alanine-substitution within core regions i, ii, iii and iv identified re ...201121262517
characterization of the porcine p65 subunit of nf-?b and its association with virus antibody levels.nf-?b p65 subunit plays important roles in controlling both innate and adaptive immunity. here we report the characterization of porcine nf-?b p65 subunit (pp65). pp65 shows high similarity to other mammalian counterparts. pp65 mrna expression was mainly observed in lung, spleen, liver and small intestine. furthermore, overexpression of pp65 activates nf-?b in porcine endothelial cell line piec, porcine alveolar macrophages cell line 3d4/21 and porcine primary fetal fibroblasts. a cooh-terminal ...201121269694
biosynthesis of classical swine fever virus nonstructural proteins.proteolytic processing of polyproteins is considered a crucial step in the life cycle of most positive-strand rna viruses. an enhancement of ns2-3 processing has been described as a major difference between the noncytopathogenic (non-cp) and the cytopathogenic (cp) biotypes of pestiviruses. the effects of accelerated versus delayed ns2-3 processing on the maturation of the other nonstructural proteins (nsp) have never been compared. in this study, we analyzed the proteolytic processing of nsp in ...201121270154
modeling of the toll-like receptor 3 and a putative toll-like receptor 3 antagonist encoded by the african swine fever virus.african swine fever virus (asfv) is a large double-stranded dna virus responsible for a lethal pig disease, to which no vaccine has ever been obtained. its genome encodes a number of proteins involved in virus survival and transmission in its hosts, in particular proteins that inhibit signaling pathways in infected macrophages and, thus, interfere with the host's innate immune response. a recently identified novel asfv viral protein (pi329l) was found to inhibit the toll-like receptor 3 (tlr3) s ...201121280117
phylogenetic analysis of the e2 gene of classical swine fever virus from the guangxi province of southern china.in this study, suspected classical swine fever (csf) samples from the guangxi province of china were obtained from pigs with acute csf, aborted fetuses, newborn pigs that died at 1-2 days of age, tonsils of healthy pigs, and leukocytes of immunized sows during 2001-2009. about 92 of 775 samples were found to be positive by rt-pcr, and 41 isolates were obtained. phylogenetic analysis was performed on the 31 isolates by sequencing the e2 gene, and the isolates were found to cluster into two groups ...201121287256
experimental infection of common warthogs (phacochoerus africanus) and bushpigs (potamochoerus larvatus) with classical swine fever virus. i: susceptibility and transmission.an incursion of classical swine fever virus (csfv) into the domestic pig population in south africa, identified in 2005, raised the concern that infection might spread to wildlife species and be maintained in these hosts. this study sought to determine whether two wildlife suidae species present in south africa, the bushpig (potamochoerus larvatus) and the common warthog (phacochoerus africanus), could support productive csfv infection. both species could be infected with csfv and transmitted in ...201121294855
influence of the 5'-proximal elements of the 5'-untranslated region of classical swine fever virus on translation and replication.the 5'-terminal sequence spanning nt 1-29 of the 5'-untranslated region of classical swine fever virus (csfv) forms a 5'-proximal stem-loop structure known as domain ia. deletions and replacement mutations were performed to examine the role of this domain. deletion of the 5'-proximal nucleotides and disruption of the stem-loop structure greatly increased internal ribosome entry site-mediated translation but abolished the replication of the replicons. internal deletions resulting in a change in t ...201121307229
dual promoter lentiviral vector generates transgenic mice expressing e2-csfv glycoprotein in their milk, but impairs early identification of transgenic embryos.lentiviral vectors containing the green fluorescent protein gene have been successfully used to select transgenic embryos before transfer to a surrogate mother. however, there are apparently no reports regarding early detection of transgenic embryos using a lentiviral vector carrying an additional transcription unit for tissue-specific expression of a valuable protein. in this study, two hiv-based lentiviral vectors were constructed. the first one contained the green fluorescent protein (gfp) co ...201121316751
classical swine fever virus ns2 protein promotes interleukin-8 expression and inhibits mg132-induced apoptosis.classical swine fever (csf) caused by virulent strains of classical swine fever virus (csfv) is a hemorrhagic disease of pigs and is characterized by disseminated intravascular coagulation, thrombocytopenia, and immunosuppression. until now, the role of the ns2 protein produced by csfv in the pathogenesis of csf is not well understood. in this report, we investigated the function of csfv ns2 by examining its effects on the pro-inflammatory cxc chemokine, interleukin-8 (il-8) expression, and cell ...201121318239
assessment of interactions between african swine fever virus, bushpigs (potamochoerus larvatus), ornithodoros ticks and domestic pigs in north-western madagascar.since its introduction in madagascar in 1998, african swine fever (asf) has severely affected national pig production and persists as a common disease in that country. two of its natural hosts in the african continent, the bushpig (potamochoerus larvatus) and tick vectors of the ornithodoros moubata complex, are reported in west and central regions of the island. however, their role in the maintenance and transmission of the virus has been insufficiently studied. in this work, we tried to assess ...201121320295
structural features of the seneca valley virus internal ribosome entry site (ires) element: a picornavirus with a pestivirus-like ires.the rna genome of seneca valley virus (svv), a recently identified picornavirus, contains an internal ribosome entry site (ires) element which has structural and functional similarity to that from classical swine fever virus (csfv) and hepatitis c virus, members of the flaviviridae. the svv ires has an absolute requirement for the presence of a short region of virus-coding sequence to allow it to function either in cells or in rabbit reticulocyte lysate. the ires activity does not require the tr ...201121325406
lack of phylogenetic evidence that the shimen strain is the parental strain of the lapinized chinese strain (c-strain) vaccine against classical swine fever.the chinese hog cholera lapinized virus (hclv), also called the "chinese strain" or "c-strain" of classical swine fever virus (csfv), was developed in china in the 1950s. there are uncertainties about the genetic heterogeneity and origin of this vaccine virus. the objectives of this study were to investigate the genetic heterogeneity of the c-strain, for which nucleotide sequences have been submitted to genbank from different laboratories, and to determine whether there is any evidence to suppor ...201121340740
detection of porcine circovirus type 1 in commercial porcine vaccines by loop-mediated isothermal amplification.a loop-mediated isothermal amplification (lamp) method with a real-time monitoring system was developed for the detection of porcine circovirus type 1 (pcv1) in commercial swine vaccines. this method was highly specific for pcv1. no cross-reaction to porcine circovirus type 2, porcine parvovirus, pseudorabies virus, classical swine fever virus, and porcine reproductive and respiratory syndrome virus was observed. the analytical sensitivity of the lamp for pcv1 dna was 10 copies/μl in the case of ...201121948286
[antigenic diversity of african swine fever viruses].data on the seroimmunotypic and hemadsorbing characteristics of african swine fever virus (asf) are summarized. according to the results of immunological sampling in pigs and those of hemagglutination inhibition test, the known asfv strains and isolates were divided into 11 groups, 8 were characterized as seroimmunogroups having their specific reference strains. a 110-140-kd asfv serotype-specific nonstructural major glycoprotein was identified. it is suggested that it is the glycoprotein that c ...201121899069
a novel tlr3 inhibitor encoded by african swine fever virus (asfv).african swine fever virus (asfv) encodes proteins that manipulate important host antiviral mechanisms. bioinformatic analysis of the asfv genome revealed orf i329l, a gene without any previous functional characterization as a possible inhibitor of tlr signaling. we demonstrate that orf i329l encodes a highly glycosylated protein expressed in the cell membrane and on its surface. i329l also inhibited dsrna-stimulated activation of nfκb and irf3, two key players in innate immunity. consistent with ...201121203785
a novel spatial and stochastic model to evaluate the within- and between-farm transmission of classical swine fever virus. i. general concepts and description of the model.a new stochastic and spatial model was developed to evaluate the potential spread of classical swine fever virus (csfv) within- and between-farms, and considering the specific farm-to-farm contact network. within-farm transmission was simulated using a modified si model. between-farm transmission was assumed to occur by direct contacts (i.e. animal movement) and indirect contacts (i.e. local spread, vehicle and person contacts) and considering the spatial location of farms. control measures dict ...201120708351
interferon status and white blood cells during infection with african swine fever virus in vivo.african swine fever virus (asfv) is the causative agent of african swine fever that is the significant disease of domestic pigs, with high rates of mortality. asfv is double-stranded dna virus whose genes encode some proteins that are implicated in the suppression of host immune response. in this study, we have modeled in vivo infection of asfv for determination of interferon (ifn) status in infected pigs. we measured the level of ifn-α, -β and -γ by enzyme-linked immunosorbent assay and showed ...201122226551
phylogenetic analysis of recent classical swine fever virus (csfv) isolates from assam, india.classical swine fever (csf), a highly contagious viral disease of pigs, is endemic in india. as there is no information concerning the accurate genetic typing of classical swine fever virus (csfv) isolates in india, 16 csf viruses isolated during 2005-2007 from domestic pigs in different districts of assam were typed in 5' utr (150 nucleotides). to confirm the genetic typing results and to study the genetic variability, selected viruses were also analyzed in e2 (190 nt) and ns5b gene (409 nt) re ...201119896713
protective efficacy of a classical swine fever virus c-strain deletion mutant and ability to differentiate infected from vaccinated animals.classical swine fever (csf) continues to be the most economically damaging pig disease in the world. the disease can be effectively controlled by vaccination with the live c-strain vaccine. this vaccine, however, does not enable the serological differentiation between infected and vaccinated animals (diva) and its use can therefore impose severe trade restrictions. csf-specific diagnostic elisas detect antibodies directed against the conserved and immunodominant a domain of the e2 structural gly ...201120541334
improved sero-monitoring assay for classical swine fever (csf) using the recombinant e2 protein of a recent korean isolate.classical swine fever (csf) is a highly contagious disease of pigs that causes fever, diarrhea and paralysis, often resulting in death. e2 is the major structural protein of the csf virus (csfv) and mediates the entrance of the virus, subsequently inducing a neutralizing immune response. in this study, the e2 gene of a recent korean isolate of csf, sw03, was cloned and the dna sequence was compared to other strains via phylogenetic analysis. with the purified e2 protein, an enzyme-linked immunos ...201120591455
transmission of classical swine fever virus depends on the clinical course of infection which is associated with high and low levels of virus excretion.infection with moderately virulent strains of classical swine fever virus (csfv) can lead to different courses of disease: either (sub)acute, resulting in death or recovery, or chronic disease. the virus excretion dynamics between these courses are quite dissimilar, but it is not known if this also results in differences in virus transmission. in this study, the excretion and transmission dynamics of the moderately virulent paderborn strain were studied in 15 one-to-one experiments. in these exp ...201120674195
peptide vaccine candidates against classical swine fever virus: t cell and neutralizing antibody responses of dendrimers displaying e2 and ns2-3 epitopes.three peptide-based systems integrating b and t antigenic sites of csfv and displaying the b epitopes in fourfold presentation have been designed and produced, and shown to bring about significant enhancements in immunogenicity over the peptides in monomeric form. of the different strategies tested for producing the dendrimeric constructs, stepwise spps using 3,6-dioxaoctanoic acid as flexible, peg-like spacer units at the branching points is clearly advantageous, in particular over ligation in ...201120812373
isolation and identification of a bovine viral diarrhea virus from sika deer in china.bovine viral diarrhea virus (bvdv) infections continue to cause significantly losses in the deer population. better isolation and identification of bvdv from sika deer may contribute significantly to the development of prophylactic therapeutic, and diagnostic reagents as well as help in prevention and control of bvdv. however, isolation and identification of bvdv from sika deer is seldom reported in literature. in this study, we collected some samples according to clinical sign of bvdv to isolat ...201121352530
genetic characterisation of african swine fever viruses from recent and historical outbreaks in sardinia (1978-2009).three discrete regions of the african swine fever virus (asfv) were analysed in the genomes of a wide range of isolates collected from wild and domestic pigs in sardinia, over a 31-year period (1978-2009). the analysis was conducted by genotyping based on sequence data from three single copy asf genes. the e183l gene encoding the structural protein p54 and part of the gene encoding the p72 protein were used to delineate genotypes, before intra-genotypic resolution of viral relationships by analy ...201121373994
proteomic analysis of swine serum following highly virulent classical swine fever virus infection.classical swine fever virus (csfv) belongs to the genus pestivirus within the family flaviviridae. virulent strains of classical swine fever virus (csfv) cause severe disease in pigs characterized by immunosuppression, thrombocytopenia and disseminated intravascular coagulation, which causes significant economic losses to the pig industry worldwide.201121385403
hax-1: a regulator of calcium signaling and apoptosis progression with multiple roles in human disease.introduction: hax-1, the hematopoietic cell-specific protein-associated protein x-1, is an inhibitor of apoptosis, which has been implicated in severe congenital neutropenia (scn), neurological disorders and cancer. hax-1 over-expression, as found in numerous types of cancer, results in resistance to granzyme b and caspase-3 and stabilizes the x-linked inhibitor of apoptosis, whereas its absence or knockdown promotes apoptosis. hax-1 is bound to the cytosolic faces of mitochondria and the endopl ...201121391832
identification of host cell binding peptide from an overlapping peptide library for inhibition of classical swine fever virus infection.the envelope proteins of classical swine fever virus (csfv) mediate the binding of csfv to cell surface molecules and allow csfv subsequent to enter host cells. however, the proteins binding to host cells and their binding sequences are uncertain. the results showed that the protein e1, e2, and erns were displayed on the surfaces of t7 phages. the e2 and erns phage clones showed high binding affinity to host cells, in which the e2 phage clone interacted more specifically with host cells than wit ...201121400206
attenuation of nitric oxide bioavailability in porcine aortic endothelial cells by classical swine fever virus.classical swine fever (csf) causes severe disease in pigs, characterized by hemorrhage, fever, and leucopenia. a primary target of the virus is endothelial cells, where a pro-inflammatory and pro-coagulant response occurs with downregulation of gap junctional communication; these changes establish a basis for haemostatic imbalance. the aim of this study was to gain an understanding of the effect of classical swine fever virus (csfv) on endothelial nitric oxide synthase (enos) expression and nitr ...201121424728
[pathology of lymphoid tissue cells infected by african swine fever virus in vitro].the authors studied the pathology of bone marrow (bm) lymphoid cell from pigs infected by african swine fever virus (asfv) in vitro. monocytes were shown to be primarily afflicted in unstimulated bm culture. these cells disappeared completely 72 hours after infection. just 24 hours following asfv infection, there were atypical lymphocytes amounting to 12% of the general lymphoid population at hour 72 after inoculation.the area and perimeter of minor, middle, and large lymphocytes tended to reduc ...201121427953
prevalence of classical swine fever virus in india: a 6-year study (2004-2010).a study was undertaken regarding the prevalence of classical swine fever virus (csfv) antibodies and antigens in sera and suspected tissue samples of domestic pigs. the samples were received between january 2004 and september 2010. a total of 594 serum samples from 12 states and 287 tissue samples from 13 states of india were tested using commercial enzyme-linked immunosorbent assay (elisa) kits. the mean prevalence of csfv antibodies in suspected sera was 63.3% (376/594), whereas 76.7% (220/287 ...201121429182
improvement of the immunity of pig to hog cholera vaccine by recombinant plasmid with porcine interleukin-6 gene and cpg motifs.in order to observe the dosage-effect of recombinant pig interleukin-6 gene and cpg motifs on the immune responses of swine to vaccine, a novel recombinant eukaryotic vpil6c plasmid was packed with chitosan nanoparticles (cnp) prepared by ionic cross linkage, which contains pig interleukin-6 gene and immunostimulatory sequence consisted of 11 cpg motifs. cnp-vril6c was then utilized to inoculate 30-day-old piglets intramuscularly at the dosage of 0.5, 1.0 and 1.5mg/per capita, respectively. mean ...201121443961
species-specific variation in rela underlies differences in nf-?b activity: a potential role in african swine fever pathogenesis.african swine fever virus (asfv) is a highly infectious disease of domestic pigs, with virulent isolates causing a rapidly fatal hemorrhagic fever. in contrast, the porcine species endogenous to africa tolerate infection. the ability of the virus to persist in one host while killing another genetically related host implies that disease severity may be, in part, modulated by host genetic variation. to complement transcription profiling approaches to identify the underlying genetic variation in th ...201121450812
genetic diversity of porcine circovirus type 2 (pcv2) in the romanian wild boar population.in this study, we have analyzed 23 pcv2 orf2 sequences recovered from wild boar population in romania. the pcv2 sequences were originated from different geographical regions in romania, and collected between 2008 and 2009 during the classical swine fever virus (csfv) surveillance campaign. complete open reading frame 2 (orf2) nucleotide sequences were obtained and compared with sequences mainly from european and asian isolates. the romanian sequences were identified as belonging to previously de ...201121453946
viewpoint: factors involved in type i interferon responses during porcine virus infections.since type i interferon (ifn-i) is considered a potent antiviral defence mechanism, it is not surprising that during evolution viruses have development of various mechanisms to counteract ifn-i induction or release. despite this, certain virus infections are associated with very high levels of systemic ifn-i. one explanation for this observation is the presence of plasmacytoid dendritic cells (pdc), which are able to produce high levels of ifn-i despite the presence of viral ifn-i antagonists. e ...201121458079
development and validation of a novel sybr green real-time rt-pcr assay for the detection of classical swine fever virus evaluated on different real-time pcr platforms.classical swine fever is a highly contagious viral disease that causes significant economic losses in pig production on a global scale. the rapid dissemination of the virus and the variability of the clinical signs merit the development of swift and accurate classical swine fever virus (csfv) detection methods, which can assist in disease control. the development and evaluation of a novel quantitative real-time rt-pcr assay for csfv detection, based on sybr green coupled to melting curve analysi ...201121458490
vaccination with viral protein-mimicking peptides postpones mortality in domestic pigs infected by african swine fever virus.periodic outbreaks of african swine fever virus (asfv) infection around the world threaten local populations of domestic pigs with lethal disease and provide grounds for pandemic spread. effective vaccination may bring this threat under control. we investigated the effectiveness of select peptides mimicking viral proteins in establishing a protective immune response. forty-six synthetic peptides based on the analysis of the complete nucleotide sequence of asfv were tested for immunogenicity in m ...201121468582
genomic analysis of highly virulent georgia 2007/1 isolate of african swine fever virus.african swine fever is widespread in africa but has occasionally been introduced into other continents. in june 2007, african swine fever was isolated in the caucasus region of the republic of georgia and subsequently in neighboring countries (armenia, azerbaijan, and 9 states of the russian federation). previous data for sequencing of 3 genes indicated that the georgia 2007/1 isolate is closely related to isolates of genotype ii, which has been identified in mozambique, madagascar, and zambia. ...201121470447
partial protection against classical swine fever virus elicited by dendrimeric vaccine-candidate peptides in domestic pigs.we report the immunogenicity of three dendrimeric peptide vaccine candidates for classical swine fever virus (csfv). each dendrimeric construct contained four copies of a b-cell epitope from the e2 glycoprotein of csfv [construct 1: e2 (694-712); 2: e2 (712-727); 3: e2 (829-842)] joined to a t-cell epitope from the ns3 protein (residues 1446-1460). intramuscular immunization of domestic pigs with the different constructs significantly reduced the clinical score after lethal challenge with csfv. ...201121496472
molecular characterization of lapinized classical swine fever vaccine strain by full-length genome sequencing and analysis.the complete genome of a lapinized classical swine fever virus (csfv) vaccine strain was amplified into nine overlapping fragments by rt-pcr, and nucleotide sequences were determined. complete genome sequence alignment and phylogenetic analysis indicated 92.6-98.6% identities at the nucleotide level with other reported csfv strains and could be grouped into subgroup 1.1 along with other attenuated strains of csfv. the 5'-utr demonstrated >97.0% nucleotide similarity with most of vaccine csfv str ...201121500112
species characterization in the genus pestivirus according to palindromic nucleotide substitutions in the 5'-untranslated region.the palindromic nucleotide substitutions (pns) at the three variable loci (v1, v2 and v3) in the 5'-untranslated region (utr) of the pestivirus genome have been considered for taxonomical segregation of the species, through the evaluation of 534 strains. on the basis of qualitative and quantitative secondary structure characteristics, species have been identified within the genus, determining genetic distances between species isolates, clarifying borderline and multirelated sequences, and charac ...201121514324
dynamic distribution and tissue tropism of classical swine fever virus in experimentally infected pigs.abstract:201121535885
development of a triplex taqman real-time rt-pcr assay for differential detection of wild-type and hclv vaccine strains of classical swine fever virus and bovine viral diarrhea virus 1.in this study, genomic sequences of pestiviruses available in genbank were aligned to design three primer pairs and taqman probes: two targeting the ns5a region of the viral genome of classical swine fever virus (csfv) for the differentiation of wild-type csfv and hog cholera lapinized vaccine (hclv) vaccine, and one targeting the 5'-untranslated region of bovine viral diarrhea virus 1 (bvdv-1). with these primers and probes, a triplex taqman real-time rt-pcr assay was developed for differentiat ...201121536312
protection of european domestic pigs from virulent african isolates of african swine fever virus by experimental immunisation.african swine fever (asf) is an acute haemorrhagic disease of domestic pigs for which there is currently no vaccine. we showed that experimental immunisation of pigs with the non-virulent ourt88/3 genotype i isolate from portugal followed by the closely related virulent ourt88/1 genotype i isolate could confer protection against challenge with virulent isolates from africa including the genotype i benin 97/1 isolate and genotype x uganda 1965 isolate. this immunisation strategy protected most pi ...201121549789
comparative inhibitory activity of the stilbenes resveratrol and oxyresveratrol on african swine fever virus replication.stilbenols are polyphenolic phytoalexins produced by plants in response to biotic or abiotic stress. these compounds have received much attention because of their significant biological effects. one of these is their antiviral action, which has previously been documented for two members of this class, namely resveratrol and oxyresveratrol. here we tested the antiviral effect of these two compounds on african swine fever virus, the only member of the newly created family asfarviridae and a seriou ...201121557969
substitution of specific cysteine residues in e1 glycoprotein of classical swine fever virus strain brescia affects formation of e1-e2 heterodimers and alters virulence in swine.e1, along with e(rns) and e2, is one of the three envelope glycoproteins of classical swine fever virus (csfv). e1 and e2 are anchored to the virus envelope at their carboxyl termini and e(rns) loosely associates with the viral envelope. in infected cells, e2 forms homodimers and heterodimers with e1 mediated by disulfide bridges between cysteine residues. the e1 protein of csfv strain brescia contains six cysteine residues at positions 5, 20, 24, 94, 123 and 171. the role of these residues in t ...201121561909
identification of two internal signal peptide sequences: critical for classical swine fever virus non-structural protein 2 to trans-localize to the endoplasmic reticulum.abstract: background: the membrane topology and molecular mechanisms for endoplasmic reticulum (er) localization of classical swine fever virus (csfv) non-structural 2 (ns2) protien is unclear. we attempted to elucidate the subcellular localization, and the molecular mechanisms responsible for the localization of this protein in our study. the ns2 gene was amplified by reverse transcription polymerase chain reaction, with the transmembrane region and hydrophilicity of the ns2 protein was predict ...201121592340
emergence of classical swine fever virus in israel in 2009.classical swine fever (csf) re-emerged in israel in february 2009 after an absence of 62years. the outbreak occurred on a domestic pig farm in northern israel and affected domestic pigs and wild boar. on the basis of phylogenetic analysis of a 190 base pair fragment of the e2 glycoprotein gene, the israeli csf virus strain belonged to genotype 2.1 and was genetically most similar to a chinese csf virus strain.201121592830
a generic real-time taqman assay for specific detection of lapinized chinese vaccines against classical swine fever.classical swine fever (csf) is a highly contagious disease, causing severe economic losses in the pig industry worldwide. vaccination of pigs with lapinized chinese vaccines is still practised in some regions of the world, where the virus is enzootic, in order to prevent and control the disease. however, a single real-time assay that can detect all lapinized chinese vaccines used widely, namely, lapinized philippines coronel (lpc), hog cholera lapinized virus (hclv) and the riems c-strain is sti ...201121600240
interactions of the dna polymerase x from african swine fever virus with the ssdna. properties of the total dna-binding site and the strong dna-binding subsite.interactions of the polymerase x from the african swine fever virus with the ssdna have been studied, using quantitative fluorescence titration and fluorescence resonance energy transfer techniques. the primary dna-binding subsite of the enzyme, independent of the dna conformation, is located on the c-terminal domain. association of the bound dna with the catalytic n-terminal domain finalizes the engagement of the total dna-binding site of the enzyme and induces a large topological change in the ...201121601347
kinetic mechanism of the ssdna recognition by the polymerase x from african swine fever virus. dynamics and energetics of intermediate formations.kinetic mechanism of the ssdna recognition by the polymerase x of african swine fever virus (asfv) and energetics of intermediate formations have been examined, using the fluorescence stopped-flow method. the association is a minimum three-step processthe nucleic acid makes the initial contact through the c-terminal domain, which generates most of the overall δg°. in the second step the nucleic acid engages the n-terminal domain, assuming the bent structure. in equilibrium, the complex exists in ...201121605932
virus inactivation by salt (nacl) and phosphate supplemented salt in a 3d collagen matrix model for natural sausage casings.due to possible presence and spread of contagious animal viruses via natural sausage casings the international trade in these food products is subject to veterinary and public health requirements. in order to manage these restrictions we determined the effect of casing preservation on four highly contagious viruses for livestock: foot-and-mouth-disease virus (fmdv), classical swine fever virus (csfv), swine vesicular disease virus (svdv) and african swine fever virus (asfv). we used an in vitro ...201121632134
genetic diversity and positive selection analysis of classical swine fever virus isolates in south china.classical swine fever virus (csfv) causes a highly contagious disease that leads to significant economic losses in the pig industry worldwide. however, there is a paucity of knowledge on the accurate genotyping of csfv isolates in south china. this study genotyped the e2 gene of 14 csfv strains isolated during 2008-2010 from domestic pigs in different districts of south china. phylogenetic analyses revealed that all of the 14 csfv isolates were clustered into genetic subgroup 1.1. this contrasts ...201121643769
the persistence of african swine fever virus in field-infected ornithodoros erraticus during the asf endemic period in portugal.african swine fever (asf) is an important disease of pigs and outbreaks of asf have occurred in europe on multiple occasions. to explore the period for which the european soft tick species ornithodoros erraticus (acari: argasidae) is able to act as a reservoir of african swine fever virus (asfv) after infected hosts are removed, we collected specimens from farms in the provinces of alentejo and algarve in portugal during the endemic period and tested them subsequently using cell culture and expe ...201121655242
effects of the polymorphisms of mx1, bat2 and cxcl12 genes on immunological traits in pigs.it is necessary that genetic markers or biomarkers can be used to predict resistance towards a wide range of infectious diseases. in the present study, we estimated the potential markers and measured their relationship with heritabilities of a wide range of immune traits. polymorphisms in exon 13 of mx1, intron 25 of bat2 and intron 3 of cxcl12 were identified by sequencing, and the genotypes were analyzed by pcr-rflp in a resource population composed of 352 pure breed landrace piglets at days 0 ...201121667240
african swine fever virus p72 genotype ix in domestic pigs, congo, 2009.african swine fever virus p72 genotype ix, associated with outbreaks in eastern africa, is cocirculating in the republic of the congo with west african genotype i. data suggest that viruses from eastern africa are moving into western africa, increasing the threat of outbreaks caused by novel viruses in this region.201121801650
multiple linear b-cell epitopes of classical swine fever virus glycoprotein e2 expressed in e.coli as multiple epitope vaccine induces a protective immune response.abstract: classical swine fever is a highly contagious disease of swine caused by classical swine fever virus,an oie list a pathogen.epitope-based vaccines is one of the current focuses in the development of new vaccines against classical swine fever virus (csfv). two b-cell linear epitopes re2-ba from the e2 glycoprotein of csfv, re2-a (cfrrekpfphrmdcvtttvened, aa 844-865) and re2-b (ckedyryaisstneigllgagglt, aa 693-716), were constructed and heterologously expressed in escherichia coli as mult ...201121801433
chemical disinfection of high-consequence transboundary animal disease viruses on nonporous surfaces.disinfection is a critical part of the response to transboundary animal disease virus (tadv) outbreaks by inactivating viruses on fomites to help control infection. to model the inactivation of tadv on fomites, we tested selected chemicals to inactivate foot and mouth disease virus (fmdv), african swine fever virus (asfv), and classical swine fever virus (csfv) dried on steel and plastic surfaces. for each of these viruses, we observed a 2 to 3 log reduction of infectivity due to drying alone. w ...201121798759
virus distribution and role of thymic macrophages during experimental infection with noncytopathogenic bovine viral diarrhea virus type 1.thymic depletion, presence of viral antigen, and changes in distribution and cytokine production of thymic macrophages were investigated in calves experimentally infected with a noncytopathogenic bovine viral diarrhea virus type (bvdv) 1 strain. ten clinically healthy colostrum-deprived calves were used. eight calves were inoculated with the virus and two were used as uninfected controls. calves were sedated and euthanized in batches between 3 and 14 days postinoculation. at necropsy, thymus sam ...201121768605
classical swine fever virus suppresses maturation and modulates functions of monocyte-derived dendritic cells without activating nuclear factor kappa b.classical swine fever virus (csfv) compromises the host immune system, causing the severe disease of pigs. dendritic cells (dcs) are the most potent inducers of immune responses. in the present study, we investigated the functional properties of porcine monocyte-derived dcs (mo-dcs) affected by csfv. results showed that the expression of surface markers of dcs such as major histocompatibility complex class ii (mhc-ii), cd80, cd83 and cd86 were unimpaired, but an obviously increased expression of ...201121764089
in vitro inhibition of csfv replication by multiple sirna expression.classical swine fever (csf) is a highly contagious viral disease of pigs which causes major economic losses worldwide. no specific drug is currently available for the effective treatment of csfv infection; however, rna interference (rnai) has been applied successfully to inhibit the replication of human and other animal viruses. in this study, three effective sirnas targeting ns3 of csfv were selected. sins3-2 targeting ns3 gene was chosen for further experimentation, while sin1 and sin2 targeti ...201121699919
quantitative risk assessment for the introduction of african swine fever virus into the european union by legal import of live pigs.the recent incursion and spread of african swine fever virus (asfv) in the russian federation and caucasus region, close to european union (eu) borders, have increased the concerns regarding the probability of asfv introduction into the eu. there are many potential routes of asfv entry into eu, but here we specifically aimed to assess the probability of asfv introduction by legal trade of pigs, which historically has been one of the most important ways of exotic diseases introduction into the eu ...201121831148
enhancing dna immunization by targeting asfv antigens to sla-ii bearing cells.one of the main criticisms to dna vaccines is the poor immunogenicity that they confer on occasions, at least in large animals. confirming this theory, immunization with plasmid dna encoding two african swine fever virus genes in frame (pcmv-pq), failed in inducing detectable immune responses in pigs, while it was successful in mice. aiming to improve the immune responses induced in swine, a new plasmid was constructed, encoding the viral genes fused in frame with a single chain variable fragmen ...201121679736
virus load in pigs affected with different clinical forms of classical swine fever.summary classical swine fever (csf) is an endemic disease in india, but the real magnitude of the problem is not known as only outbreaks of acute csf are reported and many cases of chronic and clinically inapparent forms of the disease, which manifest a confusing clinical picture, remain undiagnosed. the real status of classical swine fever virus (csfv) infection can only be known by testing pigs with highly specific and sensitive diagnostic assays. to obtain the baseline prevalence of csfv infe ...201121827642
classical swine fever virus n(pro) limits type i interferon induction in plasmacytoid dendritic cells by interacting with interferon regulatory factor 7.viruses are detected by different classes of pattern recognition receptors that lead to the activation of interferon regulatory factors (irf) and consequently to the induction of alpha/beta interferon (ifn-+¦/+¦). in turn, efficient viral strategies to escape the type i ifn-induced antiviral mechanisms have evolved. previous studies established that pestivirus n(pro) antagonizes the early innate immune response by targeting the transcription factor irf3 for proteasomal degradation. here, we repo ...201121680532
immunomodulatory effect of swine ccl20 chemokine in dna vaccination against csfv.the objective of this work was to explore whether a plasmid expressing ccl20 chemokine could improve the immune response against csfv in co-administration with a dna vaccine expressing the e2 protein. the immunization of pigs with the dna vaccine formulation, that contains swine ccl20 chemokine, resulted in the homogenous induction of detectable levels of csfv antibodies at 36 days after the first injection. remarkably, immunized animals with e2 dna vaccine in co-administration with the plasmid ...201121684019
production of classical swine fever virus envelope glycoprotein e2 as recombinant polyhedra in baculovirus-infected silkworm larvae.although, classical swine fever virus (csfv) envelope glycoprotein e2 subunit vaccine has been developed using the baculovirus expression system, the expression of viral antigens in baculovirus-infected insect cells is often ineffective. therefore, an alternative strategy to the traditional baculovirus expression system is needed that is more productive and effective. here, we report a novel strategy for the large-scale production of a csfv e2 in the larvae of a baculovirus-infected silkworm, bo ...201121706129
porcine bocaviruses: genetic analysis and prevalence in chinese swine population.summaryin members of the bocavirus genus, that contain three open reading frames (orfs) of the parvovirinae subfamily, porcine bocaviruses (pobovs) exhibit the most genetic diversity. based on the orf2-encoded viral protein (vp1) classification, the six reported porcine bocaviruses were grouped into four species: pobov1 (porcine boca-like virus or pbolv), pobov2 (porcine parvovirus 4 or ppv4), pobov3 (pbov1/pbov2) and pobov4 (6v/7v), with pobov3 and pobov4 each having two genotype viruses. all f ...201121676363
hur binding to au-rich elements present in the 3' untranslated region of classical swine fever virus.classical swine fever virus (csfv) is the member of the genus pestivirus under the family flaviviridae. the 5' untranslated region (utr) of csfv contains the ires, which is a highly structured element that recruits the translation machinery. the 3' utr is usually the recognition site of the viral replicase to initiate minus-strand rna synthesis. adenosine-uridine rich elements (are) are instability determinants present in the 3' utr of short-lived mrnas. however, the presence of ares in the 3' u ...201121729330
evaluation of a real-time rt-pcr assay using minor groove binding probe for specific detection of chinese wild-type classical swine fever virus.a one-step real-time rt-pcr assay using a minor groove binding probe was developed for the specific detection of chinese wild-type classical swine fever virus (csfv). the assay detected wild-type csfv strains representing different genotypes, but did not amplify viral rna from the hog cholera lipinized virus (hclv) vaccine-strain and other porcine viruses. the assay had a detection limit of 10 copies/reaction or 3.0 median tissue culture infective dose/reaction. in comparison to the sequencing n ...201121723883
propagation of classical swine fever virus in vitro circumventing heparan sulfate-adaptation.amplification of natural virus isolates in permanent cell lines can result in adaptation, in particular enhanced binding to heparan sulfate (hs)-containing glycosaminoglycans present on most vertebrate cells. this has been reported for several viruses, including the pestivirus classical swine fever virus (csfv), the causative agent of a highly contagious hemorrhagic disease in pigs. propagation of csfv in cell culture is essential in virus diagnostics and research. adaptation of csfv to hs-bindi ...201121703305
disruption of nuclear organization during the initial phase of african swine fever virus infection.african swine fever virus (asfv), the causative agent of one of the most devastating swine diseases, has been considered exclusively cytoplasmic, even though some authors have shown evidence of an early stage of nuclear replication. in the present study, an increment of lamin a/c phosphorylation was observed in asfv-infected cells as early as 4 h postinfection, followed by the disassembling of the lamina network close to the sites where the viral genome starts its replication. at later time poin ...201121680527
A novel dendrimeric peptide induces high level neutralizing antibodies against classical swine fever virus in rabbits.The amino acid sequence (TAVSPTTLR, 829-837aa) on the glycoprotein E2 of classical swine fever virus (CSFV) is a conserved and linear neutralizing epitope. In the present study, two peptides were constructed based the core sequence of this neutralizing epitope, the dendrimeric peptide (Th-B(4)) containing four copies of B cell epitope fused to one copy of promiscuous T helper (Th) cell epitope and the peptide Th-B containing a single copy of B cell epitope fused to one copy of Th cell epitope. T ...201122051445
rapid typing of foot-and-mouth disease serotype asia 1 by reverse transcription loop-mediated isothermal amplification.a reverse transcriptase loop-mediated isothermal amplification (rt-lamp) assay was rapidly used to detect serotype asia 1 of foot-and-mouth disease virus (fmdv) within 45 min at 61°c. all fmdv serotype asia 1 reference strains were positive by rt-lamp, while other viruses such as fmdv serotypes o, c, a and classical swine fever virus, swine vesicular disease virus, porcine reproductive and respiratory syndrome virus and japanese encephalitis virus remained negative. furthermore, fmdv sreotype as ...201122040459
Rapid and sensitive detection of PRRSV by a reverse transcription-loop-mediated isothermal amplification assay.A real-time monitoring reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the sensitive and specific detection of prototypic, prevalent North American porcine reproductive and respiratory syndrome virus (PRRSV) strains. As a higher sensitivity and specificity method than reverse transcription polymerase chain reaction (RT-PCR), the RT-LAMP method only used a turbidimeter, exhibited a detection limit corresponding to a 10(-4) dilution of template RNA ex ...201121847756
experimental infection of bama miniature pigs with a highly virulent classical swine fever virus.currently, larger domestic pigs are only animals widely used in vaccine evaluation and pathogenicity study of classical swine fever virus (csfv). this study was aimed to create an alternative animal experimental infection model of csfv.201121943299
disinfection of foot-and-mouth disease and african swine fever viruses with citric acid and sodium hypochlorite on birch wood carriers.transboundary animal disease viruses such as foot-and-mouth disease virus (fmdv) and african swine fever virus (asfv) are highly contagious and cause severe morbidity and mortality in livestock. proper disinfection during an outbreak can help prevent virus spread and will shorten the time for contaminated agriculture facilities to return to food production. wood surfaces are prevalent at these locations, but there is no standardized method for porous surface disinfection; commercial disinfectant ...201122115968
classical swine fever virus detection: results of a real-time reverse transcription polymerase chain reaction ring trial conducted in the framework of the european network of excellence for epizootic disease diagnosis and control.the current study reports on a real-time reverse transcription polymerase chain reaction (real-time rt-pcr) ring trial for the detection of classical swine fever virus (csfv) genomic rna undertaken by 10 european laboratories. all laboratories were asked to use their routine in-house real-time rt-pcr protocols and a standardized protocol commonly used by the friedrich-loeffler-institute (fli) on a panel of well-characterized samples. in general, all participants produced results within the accep ...201121908363
development of an immunochromatographic strip for rapid detection of antibodies against classical swine fever virus.the genes encoding the erns and e2 antigen epitopes of classical swine fever virus (csfv) were expressed as a chimeric protein in escherichia coli bl21 by pet expression system. the antigenicity of the expressed protein cnc2 was identified by indirect enzyme-linked immunoabsorbant assay (elisa) and immunoblot with anti-csfv antibodies. based on the cnc2 protein, an immunochromatographic strip was developed to evaluate the antibody titer of serum samples from swine vaccinated with csfv vaccine ra ...201122207080
methods for growing and titrating african swine fever virus: field and laboratory samples.growing african swine fever virus (asfv) isolates obtained mainly from the field, but also engineered in the laboratory, is a critical step for diagnosis, titration, or virus infection studies. this unit describes a set of methods and protocols to produce and titrate any asfv strain in cell cultures. the procedures include (1) basic techniques to prepare virus-sensitive target cells; (2) strategies for growth, concentration, and purification of virus stocks; and (3) the semi-quantitative (end di ...201122161547
development of a loop-mediated isothermal amplification method for rapid detection of porcine boca-like virus.the porcine boca-like virus (pbo-likev) was recently discovered in swedish pigs with post-weaning multisystemic wasting syndrome (pmws). in this study, a loop-mediated isothermal amplification (lamp) assay was developed for rapid, specific and sensitive detection of pbo-likev. a set of four primers specific for six regions of pbo-likev vp1/2 genes was designed with the online software. the reaction temperature and time were optimized to 65°c and 60min, respectively. lamp products were detected b ...201122172971
characterization of african swine fever virus caucasus isolate in european wild boars.since 2007, african swine fever has spread from the caucasus region. to learn more about the dynamics of the disease in wild boars (sus scrofa), we conducted experiments by using european wild boars. we found high virulence of caucasus isolates limited potential for establishment of endemicity.201122172247
genetic variation of border disease virus species strains.the 5´-untranslated region of pestivirus strains isolated from domestic and wild animals were analysed to determine their taxonomic status according to nucleotide changes in the secondary genomic structure using the palindromic nucleotide substitutions (pns) method. a total of 131 isolates out of 536 pestivirus strains evaluated, were clustered as border disease virus (bdv) species. the bdv strains were further divided into at least 8 genotypes or subspecies. thirty-two isolates from small rumin ...201122194225
Development and inter-laboratory validation study of an improved new real-time PCR assay with internal control for detection and laboratory diagnosis of African swine fever virus.A real-time polymerase chain reaction (PCR) assay for the rapid detection of African swine fever virus (ASFV), multiplexed for simultaneous detection of swine beta-actin as an endogenous control, has been developed and validated by four National Reference Laboratories of the European Union for African swine fever (ASF) including the European Union Reference Laboratory. Primers and a TaqMan(®) probe specific for ASFV were selected from conserved regions of the p72 gene. The limit of detection of ...201121946285
development of a porcine epidemic diarrhea virus m protein-based elisa for virus detection.a membrane (m), protein-based elisa was developed to detect porcine epidemic diarrhea virus (pedv). the m gene of pedv was expressed in escherichia coli. the purified recombinant m protein was used to immunize rabbits to generate a polyclonal antibody. immunofluorescence analysis indicated that the anti-pedv-m antibody reacted with pedv-infected cells. the antibody was utilized to develop an indirect elisa to detect pedv. other viruses, porcine transmissible gastroenteritis coronavirus, avian in ...201020882317
comparison of the protective efficacy of recombinant adenoviruses against classical swine fever.classical swine fever (csf), which is caused by classical swine fever virus (csfv), is a highly contagious and often fatal swine disease that is responsible for significant losses to the swine industry worldwide. previously, we demonstrated that pigs immunized with a recombinant adenovirus (radv-e2) expressing the e2 glycoprotein of csfv were protected against virulent csfv; however, a few pigs showed a short-term fever and occasional pathological changes. to enhance the efficacy of the vaccine, ...201020923683
investigation of a pig herd with animals seropositive for classical swine fever and where porcine circovirus-associated disease had been diagnosed.aim: to investigate the cause of classical swine fever (csf) virus-seropositive animals in a nucleus pig-breeding herd in new zealand, where porcine circovirus-associated disease had been diagnosed. case history and clinical findings: an exotic disease investigation was undertaken to exclude csf and porcine reproductive and respiratory syndrome (prrs) on a nucleus pig-breeding herd comprising approximately 300 breeding sows, 1,000 weaners, and 650 grower pigs. the herd was experiencing poor repr ...201020927176
[comparison of six detection methods for classical swine fever virus].we investigated the advantages and disadvantages of six methods to detect classical swine fever virus (csfv).201020931878
human αifn co-formulated with milk derived e2-csfv protein induce early full protection in vaccinated pigs.subunit vaccines are a suitable alternative for the control of classical swine fever. however, such vaccines have as the main drawback the relatively long period of time required to induce a protective response, which hampers their use under outbreak conditions. in this work, type i interferon is used as an immunostimulating molecule in order to increase the immunogenicity of a vaccine candidate based on the e2-csfv antigen produced in goat milk. pigs vaccinated with e2-csfv antigen co-formulate ...201020933567
a novel rt-lamp assay for rapid and simple detection of classical swine fever virus.a simple and rapid assay for the detection of classical swine fever virus (csfv) was established using reverse transcription loop-mediated isothermal amplification (rt-lamp). this study describes the amplification of the genomic rna of csfv under isothermal conditions (63 °c) within one hour, using a set of six primers (two outer primers, two inner primers and two loop primers). this rt-lamp assay showed 100-fold higher sensitivity than the standard rt-pcr method and identified eighteen addition ...201020960285
an indirect elisa of classical swine fever virus based on quadruple antigenic epitope peptide expressed in e.coli.in this study, a synthesized quadruple antigenic epitope gene region of the classical swine fever virus (csfv) e2 glycoprotein was expressed in e. coli to a obtain target protein. this target protein was used as a coating antigen to establish an indirect elisa for specifically detecting anti-csfv antibodies in serum samples from pigs. the p/n cut-off value of this assay was 1.92 by receiver operating characteristic curve (roc) analysis based on 30 negative sera and 80 positive samples. the test ...201020960287
african swine fever viruses with two different genotypes, both of which occur in domestic pigs, are associated with ticks and adult warthogs, respectively, at a single geographical site.the role of the ancestral sylvatic cycle of the african swine fever virus (asfv) is not well understood in the endemic areas of eastern africa. we therefore analysed the asf infection status on samples collected from 51 free-ranging warthogs (phacocherus africanus) and 1576 ornithodorus porcinus ticks from 26 independent warthog burrows at a single ranch in kenya. abattoir samples from 83 domestic pigs without clinical symptoms, originating from specific locations with no recent reported asf out ...201020965989
small peptide inhibitors disrupt a high-affinity interaction between cytoplasmic dynein and a viral cargo protein.several viruses target the microtubular motor system in early stages of the viral life cycle. african swine fever virus (asfv) protein p54 hijacks the microtubule-dependent transport by interaction with a dynein light chain (dynll1/dlc8). this was shown to be a high-affinity interaction, and the residues gradually disappearing were mapped on dlc8 to define a putative p54 binding surface by nuclear magnetic resonance (nmr) spectroscopy. the potential of short peptides targeting the binding domain ...201020686048
in vitro inhibition of csfv replication by retroviral vector-mediated rna interference.classical swine fever (csf) is a highly contagious viral disease of pigs which causes major economic losses worldwide. no specific drug is currently available for the effective treatment of csfv infection. rna interference (rnai) technology depends on effective delivery systems, for which several effective vectors have recently been developed. three retroviral plasmids containing sirna genes targeting different regions of n(pro) and ns4a have been constructed, and 3 replication-incompetent retro ...201020691206
loop-mediated isothermal amplification for rapid detection and differentiation of wild-type pseudorabies and gene-deleted virus vaccines.a loop-mediated isothermal amplification (lamp) assay was developed specifically for detection and differentiation of pseudorabies virus (prv). one group of primers was designed to detect wild-type strains (i.e., strains with the ge gene) and the other group of primers was designed to detect both prv ge-vaccine and wild-type strains (i.e., strains with the gg gene and with or without the ge gene). after amplification by bst enzyme at a constant temperature of 65 degrees c, a laddering of bright ...201020691214
establishment of a multiplex rt-pcr assay to detect different lineages of swine h1 and h3 influenza a viruses.classical swine h1n1, emerging european avian-like h1n1 and human-like h3n2 lineages are co-circulating in the swine population in china. the reverse transcriptase polymerase chain reaction (rt-pcr) assay is an effective method for use in influenza surveillance. in this study, a multiplex rt-pcr method was developed for simultaneous identification of hemagglutinin (ha) genes derived from the three lineages of swine influenza viruses. three primer sets were designed and aimed specifically at ha g ...201020700759
characterization of essential domains and plasticity of the classical swine fever virus core protein.pestiviruses are pathogens of cloven-hoofed animals, belonging to the flaviviridae. the pestiviral particle consists of a lipid membrane containing the three envelope glycoproteins erns, e1, and e2 and a nucleocapsid of unknown symmetry, which is composed of the core protein and the viral positive-sense rna genome. the positively charged pestiviral core protein consists of 86 to 89 amino acids. to analyze the organization of essential domains, n- and c-terminal truncations, as well as internal d ...201020702631
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