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a comparative analysis on the synonymous codon usage pattern in viral functional genes and their translational initiation region of asfv.the synonymous codon usage pattern of african swine fever virus (asfv), the similarity degree of the synonymous codon usage between this virus and some organisms and the synonymous codon usage bias for the translation initiation region of viral functional genes in the whole genome of asfv have been investigated by some simply statistical analyses. although both gc12% (the gc content at the first and second codon positions) and gc3% (the gc content at the third codon position) of viral functional ...201323161403
antibody-mediated neutralization of african swine fever virus: myths and facts.almost all viruses can be neutralized by antibodies. however, there is some controversy about antibody-mediated neutralization of african swine fever virus (asfv) with sera from convalescent pigs and about the protective relevance of antibodies in experimentally vaccinated pigs. at present, there is no vaccine available for this highly lethal and economically relevant virus and all classical attempts to generate a vaccine have been unsuccessful. this failure has been attributed, in part, to what ...201323159730
african swine fever virus controls the host transcription and cellular machinery of protein synthesis.throughout a viral infection, the infected cell reprograms the gene expression pattern in order to establish a satisfactory antiviral response. african swine fever virus (asfv), like other complex dna viruses, sets up a number of strategies to evade the host's defense systems, such as apoptosis, inflammation and immune responses. the capability of the virus to persist in its natural hosts and in domestic pigs, which recover from infection with less virulent isolates, suggests that the virus disp ...201323154157
african swine fever virus replication and genomics.african swine fever virus (asfv) is a large icosahedral dna virus which replicates predominantly in the cytoplasm of infected cells. the asfv double-stranded dna genome varies in length from about 170 to 193 kbp depending on the isolate and contains between 150 and 167 open reading frames. these are closely spaced and read from both dna strands. the virus genome termini are covalently closed by imperfectly base-paired hairpin loops that are present in two forms that are complimentary and inverte ...201323142553
african swine fever virus eradication in africa.african swine fever was reported in domestic pigs in 26 african countries during the period 2009-2011. the virus exists in an ancient sylvatic cycle between warthogs (phacochoerus africanus) and argasid ticks of the ornithodoros moubata complex in many of the countries reporting outbreaks and in two further countries in the region. eradication of the virus from the countries in eastern and southern africa where the classic sylvatic cycle occurs is clearly not an option. however, the virus has be ...201323142552
virological diagnosis of african swine fever--comparative study of available tests.the rapid and reliable detection of african swine fever virus (asfv) is essential both for timely implementation of control measures to prevent the spread of disease, and to differentiate african swine fever (asf) from other pig disease with similar clinical presentations. many virological tests are currently available for the detection of asfv (live virus), antigen and genome, including virus isolation, elisa, fluorescent antibody, polymerase chain reaction (pcr) and isothermal assays. in recen ...201323131492
african swine fever virus serodiagnosis: a general review with a focus on the analyses of african serum samples.african swine fever (asf) is an infectious disease that causes heavy mortality in domestic pigs. at present there is no vaccine against asf, and eradication in countries where the disease is endemic is based only on competent diagnosis programs and the sacrifice of infected animals. due to the presence of natural attenuated strains, certain infection conditions may result in reduced mortality. in these situations, the disease can be diagnosed by detection of specific antibodies. the use of class ...201323131491
epidemiology of african swine fever virus.african swine fever virus used to occur primarily in africa. there had been occasional incursions into europe or america which apart from the endemic situation on the island of sardinia always had been successfully controlled. but following an introduction of the virus in 2007, it now has expanded its geographical distribution into caucasus and eastern europe where it has not been controlled, to date. african swine fever affects domestic and wild pig species, and can involve tick vectors. the ab ...201323123296
african swine fever virus infection in ornithodoros ticks.african swine fever virus (asfv) is an arbovirus which is vectored by soft ticks of the ornithodoros spp. and in the sylvatic cycle infects wart hogs and bush pigs. asfv infection of domestic swine causes a high mortality disease. on the other hand, asfv infection of the tick can result in a high-titered and persistent infection depending upon the asfv isolate and the tick combination. recently, morphological, classical virology (titration) and recombinant asfv have been used to study the cellul ...201323085123
african swine fever virus morphogenesis.this review summarizes recent structural and molecular biology studies related to the morphogenesis of african swine fever virus (asfv). asfv possesses icosahedral morphology and is constituted by four concentric layers: the central nucleoid, the core shell, the inner envelope and the icosahedral capsid. the extracellular virus acquires an external envelope by budding through the plasma membrane. the genes coding for 19 of the 54 structural proteins of the asfv particle are known and the localiz ...201323059353
development of a suspension microarray for the genotyping of african swine fever virus targeting the snps in the c-terminal end of the p72 gene region of the genome.african swine fever virus (asfv) causes one of the most dreaded transboundary animal diseases (tads) in suidae. african swine fever (asf) often causes high rates of morbidity and mortality, which can reach 100% in domestic swine. to date, serological diagnosis has the drawback of not being able to differentiate variants of this virus. previous studies have identified the 22 genotypes based on sequence variation in the c-terminal region of the p72 gene, which has become the standard for categoriz ...201322776009
prevalence of african swine fever virus in apparently healthy domestic pigs in uganda.african swine fever (asf) is a contagious viral disease which can cause up to 100% mortality among domestic pigs leading to serious socio-economic impact on people's livelihoods. asf is endemic in uganda and there is paucity of information on the epidemiology of the disease. the major aim of this study was to determine the seroprevalence and prevalence of african swine fever virus (asfv) in apparently healthy slaughter pigs at wambizi slaughterhouse in kampala city, uganda. we also estimated the ...201324369729
how dna polymerase x preferentially accommodates incoming datp opposite 8-oxoguanine on the template.the modified base 8-oxo-7,8-dihydro-2'-deoxyguanosine (oxog) is a common dna adduct produced by the oxidation of dna by reactive oxygen species. kinetic data reveal that dna polymerase x (pol x) from the african swine fever virus incorporates adenine (datp) opposite to oxog with higher efficiency than the non-damaged g:c basepair. to help interpret the kinetic data, we perform molecular dynamics simulations of pol x/dna complexes, in which the template base opposite to the incoming dntp (dctp, d ...201324314086
pathogenesis of highly virulent african swine fever virus in domestic pigs exposed via intraoropharyngeal, intranasopharyngeal, and intramuscular inoculation, and by direct contact with infected pigs.to investigate the pathogenesis of african swine fever virus (asfv), domestic pigs (n=18) were challenged with a range (10(2)-10(6) 50% hemadsorbing doses (had50)) of the highly virulent asfv-malawi strain by inoculation via the intraoropharyngeal (iop), intranasopharyngeal (inp), or intramuscular (im) routes. a subsequent contact challenge experiment was performed in which six iop-inoculated donor pigs were allowed to have direct contact (dc) with six naïve pigs for exposure times that varied f ...201324076499
african swine fever virus: on the move and dangerous. 201324038277
comprehensive phylogenetic reconstructions of african swine fever virus: proposal for a new classification and molecular dating of the virus.african swine fever (asf) is a highly lethal disease of domestic pigs caused by the only known dna arbovirus. it was first described in kenya in 1921 and since then many isolates have been collected worldwide. however, although several phylogenetic studies have been carried out to understand the relationships between the isolates, no molecular dating analyses have been achieved so far. in this paper, comprehensive phylogenetic reconstructions were made using newly generated, publicly available s ...201323936068
development and validation of a multiplex, real-time rt pcr assay for the simultaneous detection of classical and african swine fever viruses.a single-step, multiplex, real-time polymerase chain reaction (rt-pcr) was developed for the simultaneous and differential laboratory diagnosis of classical swine fever virus (csfv) and african swine fever virus (asfv) alongside an exogenous internal control rna (ic-rna). combining a single extraction methodology and primer and probe sets for detection of the three target nucleic acids csfv, asfv and ic-rna, had no effect on the analytical sensitivity of the assay and the new triplex rt-pcr was ...201323923045
a nanoparticle-assisted pcr assay to improve the sensitivity for rapid detection and differentiation of wild-type pseudorabies virus and gene-deleted vaccine strains.nanoparticle-assisted polymerase chain reaction (nanopcr) is a novel method for the rapid amplification of dna and has been adopted for the detection of virus because of its simplicity, rapidity, and specificity. a nanopcr assay was developed to detect and differentiate wild-type and gene-deleted pseudorabies virus (prv). three pairs of primers for nanopcr developed in this study were selected from conserved regions of prv, producing specific amplicons of 431 bp (gb), 316 bp (ge), and 202 bp (gg ...201323872268
involvement of the reparative dna polymerase pol x of african swine fever virus in the maintenance of viral genome stability in vivo.the function of the african swine fever virus (asfv) reparative dna polymerase, pol x, was investigated in the context of virus infection. pol x is a late structural protein that localizes at cytoplasmic viral factories during dna replication. using an asfv deletion mutant lacking the pol x gene, we have shown that pol x is not required for virus growth in vero cells or swine macrophages under one-step growth conditions. however, at a low multiplicity of infection, when multiple rounds of replic ...201323824796
phytochemical analysis and in-vitro anti-african swine fever virus activity of extracts and fractions of ancistrocladus uncinatus, hutch and dalziel (ancistrocladaceae).african swine fever (asf), a highly contagious fatal acute haemorrhagic viral disease of pigs currently has no treatment or vaccination protocol and it threatens the pig industry worldwide. recent outbreaks were managed by farmers with ethnoveterinary preparations with various claims of effectiveness.201323777548
prospects for development of african swine fever virus vaccines.african swine fever virus is a large dna virus which can cause an acute haemorrhagic fever in pigs resulting in high mortality. no vaccine is available, limiting options for control. the virus encodes up to 165 genes and virus particles are multi-layered and contain more than 50 proteins. pigs immunised with natural low virulence isolates or attenuated viruses produced by passage in tissue culture and by targeted gene deletions can be protected against challenge with virulent viruses. cd8+ cells ...201323689892
transmission rate of african swine fever virus under experimental conditions.african swine fever (asf) is a highly lethal, viral disease of swine. no vaccine is available, so controlling an asf outbreak is highly dependent on zoosanitary measures, such as stamping out infected herds and quarantining of affected areas. information on asf transmission parameters could allow for more efficient application of outbreak control measures. three transmission experiments were carried out to estimate the transmission parameters of two asf virus isolates: malta'78 (in two doses) an ...201323664069
antigen targeting to apc: from mice to veterinary species.antigen delivery to receptors expressed on antigen presenting cells (apc) has shown to improve immunogenicity of vaccines in mice. an enhancement of cytotoxic t lymphocyte (ctl), helper t cell or humoral responses was obtained depending on the type of apc and the surface molecule targeted. although this strategy is being also evaluated in livestock animals with promising results, some discrepancies have been found between species and pathogens. the genetic diversity of livestock animals, the dif ...201323648645
quantification of airborne african swine fever virus after experimental infection.knowledge on african swine fever (asf) transmission routes can be useful when designing control measures against the spread of asf virus (asfv). few studies have focused on the airborne transmission route, and until now no data has been available on quantities of asf virus (asfv) in the air. our aim was to validate an air sampling technique for asf virus (asfv) that could be used to detect and quantify virus excreted in the air after experimental infection of pigs. in an animal experiment with t ...201323608475
comparative evaluation of novel african swine fever virus (asf) antibody detection techniques derived from specific asf viral genotypes with the oie internationally prescribed serological tests.the presence of antibodies against african swine fever (asf), a complex fatal notifiable oie disease of swine, is always indicative of previous infection, since there is no vaccine that is currently used in the field. the early appearance and subsequent long-term persistence of antibodies combined with cost-effectiveness make antibody detection techniques essential in control programmes. recent reports appear to indicate that the serological tests recommended by the oie for asf monitoring are mu ...201322944073
laboratory methods to study african swine fever virus.we summarize findings of comparative studies in different cells cultures susceptible to asfv infection, through the analysis of virus components and infectious virus particles production, as alternative means to grow field and laboratory asfv strains. we also provide different methods to assay the infectivity of asfv samples and to purify the infective virus particles. finally we describe the general strategy to construct virus deletion mutants that can be engineered to obtain attenuated asfv st ...201323041357
african swine fever virus transcription.african swine fever virus (asfv), a large, enveloped, icosahedral dsdna virus, is currently the only known dna-containing arbovirus and the only recognized member of the family asfarviridae. its genome encodes more than 150 open reading frames that are densely distributed, separated by short intergenic regions. asfv gene expression follows a complex temporal programming. four classes of mrnas have been identified by its distinctive accumulation kinetics. gene transcription is coordinated with dn ...201323041356
molecular diagnosis of african swine fever by a new real-time pcr using universal probe library.a highly sensitive and specific real-time pcr method was developed for the reliable and rapid detection of african swine fever virus (asfv). the method uses a commercial universal probe library (upl) probe combined with a specifically designed primer set to amplify an asfv dna fragment within the vp72 coding genome region. the detection range of the optimized upl pcr technique was confirmed by analysis of a large panel (n = 46) of asfv isolates, belonging to 19 of the 22 viral p72 genotypes desc ...201322394449
preliminary mapping of non-conserved epitopes on envelope glycoprotein e2 of bovine viral diarrhea virus type 1 and 2.bovine viral diarrhea virus (bvdv) belongs together with classical swine fever virus (csfv) and border disease virus (bdv) to the genus pestivirus in the flaviviridae family. bvdv has been subdivided into two different species, bvdv1 and bvdv2 based on phylogenetic analysis. subsequent characterization of both strains revealed major antigenic differences. because the envelope glycoprotein e2 is the most immunodominant protein for all pestiviruses, the present study focused on epitope mapping by ...201323838147
numerical taxonomy of the genus pestivirus: new software for genotyping based on the palindromic nucleotide substitutions method.the genus pestivirus from the family flaviviridae is represented by four established species; bovine viral diarrhea virus 1 (bvdv-1); bovine viral diarrhea virus 2 (bvdv-2); border disease virus (bdv); and classical swine fever virus (csfv); as well a tentative species from a giraffe. the palindromic nucleotide substitutions (pns) in the 5' untranslated region (utr) of pestivirus rna has been described as a new, simple and practical method for genotyping. new software is described, also named pn ...201323684846
identification and characterization of mimotopes of classical swine fever virus e2 glycoprotein using specific anti-e2 monoclonal antibodies.classical swine fever virus (csfv) shares high nucleic acid and amino acid sequence homology with the other members of the pestivirus genus, namely bovine viral diarrhea virus and border disease virus. all three viruses are able to infect swine and generate cross reactive antibodies, which is problematic during differential diagnosis for classical swine fever (csf). toward the development of a new generation of csf specific diagnostic tools, monoclonal antibodies specific for csfv were mapped us ...201323583685
hobi-like viruses: an emerging group of pestiviruses.the genus pestivirus is composed of 4 important pathogens of livestock: bovine viral diarrhea virus 1 and 2 (bvdv-1 and bvdv-2), classical swine fever virus (csfv), and border disease virus of sheep (bdv). bvdv are major pathogens of cattle, and infection results in significant economic loss worldwide. a new putative pestivirus species, tentatively called "hobi-like," "bvdv-3," or "atypical pestiviruses," was first identified in europe in fetal bovine serum (fbs) imported from brazil. hobi-like ...201323345268
bovine viral diarrhea virus (bvdv) infections in pigs.cattle are the natural hosts of bovine viral diarrhea virus (bvdv), which causes mucosal disease, respiratory and gastrointestinal tract infections, and reproductive problems in cattle. however, bvdv can also infect goats, sheep, deer, and pigs. the prevalence of bvdv infection in pig herds has substantially increased in the last several years, causing increased economic losses to the global pig breeding industry. this article is a summary of bvdv infections in pigs, including a historical overv ...201323587625
intra-host variation structure of classical swine fever virus ns5b in relation to antiviral therapy.classical swine fever (csf) is one of most important diseases of the suidea with severe social economic consequences in case of outbreaks. antivirals have been demonstrated, in recent publications, to be an interesting alternative method of fighting the disease. however, classical swine fever virus is an rna virus which presents a challenge as intra-host variation and the error prone rna dependent rna polymerase (rdrp) could lead to the emergence/selection of resistant variants hampering further ...201323511203
two newly developed e(rns)-based elisas allow the differentiation of classical swine fever virus-infected from marker-vaccinated animals and the discrimination of pestivirus antibodies.new generations of classical swine fever virus (csfv) marker vaccines have recently been developed in order to make emergency vaccination in case of a csf outbreak more feasible. however, the application of a marker vaccine is dependent on the availability of an accompanying discriminatory test allowing differentiation of infected from vaccinated animals (diva). cp7_e2alf, the most promising live marker vaccine candidate currently available, is a genetically modified bovine viral diarrhea virus ...201322902189
genome and molecular characterization of a csfv strain isolated from a csf outbreak in south china.in the present study, the full-length nucleotide sequences of the csfv-gz-2009 strain of classical swine fever virus (csfv) isolated from a hog pen in guangdong province in china was determined. results demonstrated that the genome of csfv-gz-2009 is 12,298 nucleotides (nt) in length, is composed of a 373-nt 5'-untranslated region (utr), has an 11,697-nt open reading frame encoding a polyprotein of 3,898 amino acids, and has a 228-nt 3'-utr. genome comparison of the csfv-gz-2009 isolate (genbank ...201323296074
real-time spr characterization of the interactions between multi-epitope proteins and antibodies against classical swine fever virus.the envelope glycoprotein e2 is the major immunodominant protein of the classical swine fever virus and can induce neutralizing antibodies and protective host-immune responses in infected swine. we designed, expressed, and purified multi-epitope protein (gst-bt22) that contains a tandem repeat of the e2 antigenic-determinant residues 693-704, 770-780, and 826-843, each of which is separated by a ggssgg sequence. in the same manner, we also designed, expressed, and purified a second protein (gst- ...201323291170
the feasibility of a novel bioreactor for vaccine production of classical swine fever virus.the performance of a new type of tide mode culture system was investigated in this study. this novel bioreactor provides two separated stages, liquid and gas, for cell growth requirements. the immobilized cells absorbed the nutrient from medium during the liquid stage and subsequently were exposed directly to fresh air to absorb oxygen during the gas stage. operating with pk15 cells under optimal conditions, we obtained 2.3×10(9) cells in 500ml reactor. it is 30 times higher than the initial ino ...201323261041
infection of mouse bone marrow-derived immature dendritic cells with classical swine fever virus c-strain promotes cells maturation and lymphocyte proliferation.in this study, the interactions of classical swine fever virus (csfv) c-strain and the virulent gslz strain with mouse bone marrow-derived immature dendritic cells (bm-imdcs) were investigated for the first time. both the c-strain and the virulent gslz strain could effectively infect and replicate in mouse bm-imdcs. c-strain-infected bm-imdcs showed a greatly enhanced degree of maturation, and could effectively promote the expansion and proliferation of allogeneic naive t cells. the c-strain ind ...201324169497
classical swine fever virus infection modulates serum levels of inf-α, il-8 and tnf-α in 6-month-old pigs.several studies have highlighted the important role of cytokines in disease development of classical swine fever virus (csfv) infection. in the present study, we examined the kinetics of 7 porcine cytokines in serum from pigs infected with 3 different csfv strains. based on the clinical picture in 6-month-old danish pigs, the strains used for inoculation were classified as being of low (bergen), low to moderate (eystrup) and moderate to high (lithuania) virulence. the cytokines interferon-alpha ...201324148871
the structure of classical swine fever virus n(pro): a novel cysteine autoprotease and zinc-binding protein involved in subversion of type i interferon induction.pestiviruses express their genome as a single polypeptide that is subsequently cleaved into individual proteins by host- and virus-encoded proteases. the pestivirus n-terminal protease (n(pro)) is a cysteine autoprotease that cleaves between its own c-terminus and the n-terminus of the core protein. due to its unique sequence and catalytic site, it forms its own cysteine protease family c53. after self-cleavage, n(pro) is no longer active as a protease. the released n(pro) suppresses the inducti ...201324146623
[progress in new-type vaccines against classical swine fever].classical swine fever (csf), an acute and highly contagious disease of swine, is caused by classical swine fever virus. csf is one of the most devastating diseases to the pig industry worldwide and results in serious economic losses. currently prophylactic vaccination is still an important strategy for the control of csf. live attenuated vaccines (such as c-strain) are safe and effective. however, there are significant changes in the clinical features of csf, displaying concurrent typical and at ...201324195355
hepatitis-c-virus-like internal ribosome entry sites displace eif3 to gain access to the 40s subunit.hepatitis c virus (hcv) and classical swine fever virus (csfv) messenger rnas contain related (hcv-like) internal ribosome entry sites (iress) that promote 5'-end independent initiation of translation, requiring only a subset of the eukaryotic initiation factors (eifs) needed for canonical initiation on cellular mrnas. initiation on hcv-like iress relies on their specific interaction with the 40s subunit, which places the initiation codon into the p site, where it directly base-pairs with eif2-b ...201324185006
rna helicase is involved in the expression and replication of classical swine fever virus and interacts with untranslated region.to investigate whether cytoplasmic rna helicase a (rha) influences the expression and replication of classical swine fever virus (csfv), an sirna molecule targeted to rha was transfected into pk-15 cells. the sirna was found to reduce cytoplasmic rha. in csfv subgenomic replicon transfected cells, incubation with the sirnas negatively impacted viral ns3 and rna production. in the csfv infected cells, treatment with the sirna resulted in a significant reduction of viral replication by 65-70%. fur ...201323220337
efficient and stable rescue of classical swine fever virus from cloned cdna using an rna polymerase ii system.conventional reverse genetics for classical swine fever virus (csfv) is based on the transfection of permissive cells with either in vitro or intracellularly synthesized rna transcripts from a viral genomic cdna clone. these strategies are complicated, inefficient and time-consuming. this study is aimed to develop an improved reverse genetics method for the direct, rapid and efficient recovery of csfv from cloned cdna. the cdna clone pbrcism was constructed, which harbors the full-length genomic ...201323192173
comprehensive evaluation of the adenovirus/alphavirus-replicon chimeric vector-based vaccine radv-sfv-e2 against classical swine fever.classical swine fever (csf) is an economically important, highly contagious swine disease caused by classical swine fever virus (csfv). marker vaccines and companion serological diagnostic tests are thought to be a promising strategy for future control and eradication of csf. previously, we have demonstrated that an adenovirus-vectored semliki forest virus replicon construct expressing the e2 glycoprotein from csfv, radv-sfv-e2, induced sterile immunity against a lethal csfv challenge. in this s ...201323153441
the survey of porcine teschoviruses in field samples in china with a universal rapid probe real-time rt-pcr assay.a real-time reverse transcription polymerase chain reaction (rt-pcr) based on taqman was established and evaluated for quantitative detection of porcine teschoviruses (ptvs). a pair of primers and a taqman probe targeting on the highly conserved sequence of the 5'-untranslated region (5'-utr) of one to 11 serotypes of ptv were designed. standard plasmid dna containing pcr amplification of the 5'-utr were constructed and used to develop the real-time rt-pcr. the results indicated that the real-ti ...201323138413
co-expression of erns and e2 genes of classical swine fever virus by replication-defective recombinant adenovirus completely protects pigs against virulent challenge with classical swine fever virus.the objective of this study was to construct a recombinant adenovirus for future csfv vaccines used in the pig industry for the reduction of losses involved in csf outbreaks. the erns and e2 genes of classical swine fever virus (csfv), which encode the two main protective glycoproteins from the "shimen" strain of csfv, were combined and inserted into the replication-defective human adenovirus type-5 and named the rad-erns-e2. nine pigs were randomly assigned to three treatment groups (three pigs ...201323092714
prevalence of classical swine fever virus in domestic pigs in south korea: 1999-2011.the major policy for eradication of classical swine fever (csf) in south korea has focused on the implementation of compulsory vaccination of the susceptible pig population. a vaccine strain of csf virus, the lom strain, is used to maintain high herd seroconversion, a practice complementary to the 'stamping-out policy' and restriction of animal movement during disease outbreaks. to survey for the prevalence of csf in domestic pigs in south korea over the past 13 years (1999-2011), we tested 4 19 ...201322925439
classical swine fever virus isolates from cuba form a new subgenotype 1.4.identification and classification of classical swine fever virus (csfv) on the basis of nucleotide sequencing and phylogenetic analysis have become an important tool to study the epidemiology and to control csf disease. according to phylogenetic analyses of short sequences from the 5'nontranslated region (150 nt) and the e2 (190 nt), most csfv isolates from south and central america have been assorted to the subgenotypes 1.1 and 1.3, while csfv isolates from cuba have been allocated to subgenoty ...201322902191
future directions for the development of chlamydomonas-based vaccines.besides serving as a valuable model in biological sciences, chamydomonas reinhardtii has been used during the last decade in the biotechnology arena to establish models for the low cost production of vaccines. antigens from various pathogens including plasmodium falciparum, foot and mouth disease virus, staphylococcus aureus, classical swine fever virus (csfv) as well as some auto-antigens, have been produced in c. reinhardtii. although some of them have been functionally characterized with prom ...201324053395
a one-step rt-pcr assay to detect and discriminate porcine reproductive and respiratory syndrome viruses in clinical specimens.outbreaks of highly pathogenic porcine reproductive and respiratory syndrome virus (hp-prrsv) have led to large economic losses and, subsequently, have drawn great attention to its diagnosis and prevention. to facilitate rapid discrimination of hp-prrsv from classical prrsv (c-prrsv), we developed a one-step rt-pcr assay. primer specificities were evaluated with rna extracted from 8 viral strains and our results revealed that the primers had a high specificity for prrsv. the assay sensitivity wa ...201324035936
classical swine fever virus triggers rig-i and mda5-dependent signaling pathway to irf-3 and nf-κb activation to promote secretion of interferon and inflammatory cytokines in porcine alveolar macrophages.classical swine fever (csf) caused by csf virus (csfv) is a highly contagious disease of pigs. the rna helicases retinoic acid-inducible gene i (rig-i) and melanoma differentiation-associated gene 5 (mda-5) are differentially involved in the detection of various rna viruses. in present study, we investigated the roles of rig-i and mda-5 in eliciting antiviral and inflammatory responses to csfv shimen strain in porcine alveolar macrophages (pams).201324034559
application of real-time quantitative polymerase chain reaction to monitoring infection of classic swine fever virus and determining optimal harvest time in large-scale production.due to the non-cytopathogenic replication of classical swine fever virus (csfv) in cell culture, large-scale production of csfv using bioreactor system remains the problem of monitoring the time of maximum virus production for optimal harvest. in this study, we proposed the application of real-time quantitative pcr assay to monitoring the progress of csfv infection and yield determination in large scale. the region of ns5b of csfv responsible for csfv genome replication was used for the designat ...201324021314
detection and differentiation of classical swine fever virus strains c and shimen by high-resolution melt analysis.differentiation of classical swine fever virus (csfv) strains is crucial for the development of effective vaccination programs and in epidemiological investigations. most of current detection methods do not discriminate between wild-type csfv strains and those used in vaccines. in this study, method involving high-resolution melt (hrm) curve analysis for the simultaneous detection and differentiation of the c and shimen strains of csfv was developed. a specific fragment of the ns2 gene was ampli ...201323994467
a portable reverse transcription recombinase polymerase amplification assay for rapid detection of foot-and-mouth disease virus.foot-and-mouth disease (fmd) is a trans-boundary viral disease of livestock, which causes huge economic losses and constitutes a serious infectious threat for livestock farming worldwide. early diagnosis of fmd helps to diminish its impact by adequate outbreak management. in this study, we describe the development of a real-time reverse transcription recombinase polymerase amplification (rt-rpa) assay for the detection of fmd virus (fmdv). the fmdv rt-rpa design targeted the 3d gene of fmdv and ...201323977101
characterization of the swine u6 promoter for short hairpin rna expression and its application to inhibition of virus replication.expression of short hairpin rnas (shrnas) by the rna polymerase type iii u6 promoter is an effective and widely used strategy for rna interference (rnai) which is a sequence-specific gene silencing mechanism. the u6 promoters from human, mouse, and swine were cloned, respectively for constructing various shrna expression vectors. the transcription efficiency of each u6 promoter was analyzed for its activity to drive expression of shrna targeting enhanced green fluorescent protein (egfp) mrna in ...201323916945
anti-influenza a virus activity of uridine derivatives of 2-deoxy sugars.influenza viruses are important pathogens that cause respiratory infections in humans and animals. apart from vaccinations, antiviral drugs play a significant role in controlling spread of the disease. influenza a virus contains two membrane glycoproteins on the external part of viral envelope: hemagglutinin (ha) and neuraminidase (na), which are crucial for productive infection in target cells. in the present work, two derivatives of tunicamycin - uridine derivatives of 2-deoxy sugars (designat ...201323911991
molecular tracing of classical swine fever viruses isolated from wild boars and pigs in france from 2002 to 2011.there were three outbreaks of classical swine fever (csf) in north-eastern france between 2002 and 2011. the first two occurred in april 2002 in the moselle department, in a wild boar and pig herd, respectively, while the third occurred in april 2003, in the bas-rhin department, in a wild boar. a survey was subsequently implemented in wild boar and domestic pig populations, during which 43 csf viruses (csfvs) were genetically characterized to provide information on virus sources, trace virus evo ...201323891170
development of a loop-mediated isothermal amplification assay for the detection of porcine hokovirus.hokoviruses have recently been detected as pathogens belonging to the family parvoviridae, which comprises porcine hokovirus (phov) and bovine hokovirus (bhov). in this study, we developed a loop-mediated isothermal amplification (lamp) assay for the rapid, specific and sensitive detection of phov. a set of four primers specific for six regions within the phov vp1/2 genes was designed using online software. the reaction temperature and time were optimized at 65°c and 60 min, respectively. lamp p ...201323850717
molecular modeling and pharmacophore elucidation study of the classical swine fever virus helicase as a promising pharmacological target.the classical swine fever virus (csfv) is a major pathogen of livestock and belongs to the flaviviridae viral family. even though there aren't any verified zoonosis cases yet, the outcomes of csfv epidemics have been devastating to local communities. in an effort to shed light to the molecular mechanisms underlying the structural and drug design potential of the viral helicase, the three dimensional structure of csfv helicase has been modeled using conventional homology modeling techniques and t ...201323781407
rapid detection of foot-and-mouth disease virus, influenza a virus and classical swine fever virus by high-speed real-time rt-pcr.high sensitivity, minor risk of cross-contamination and in particular the rapid reaction time make quantitative real-time polymerase chain reaction (qpcr) assays well suited for outbreak investigations as well as for monitoring epidemics of pathogens. in this study qpcr assays for three highly contagious animal diseases, namely foot-and-mouth-disease (fmd), influenza a (ia) and classical swine fever (csf) have been developed. furthermore, an amplification control targeting 18s ribosomal rna was ...201323702025
simplified serum neutralization test based on enhanced green fluorescent protein-tagged classical swine fever virus.the neutralization immunofluorescence test (nift), currently used for detecting neutralizing antibodies (nabs) against classical swine fever virus (csfv), is time-consuming. here, a simplified neutralization test based on enhanced green fluorescent protein (egfp)-tagged csfv (egfp-nt) was developed for direct detection of anti-csfv nabs without immunostaining. the relative sensitivity and specificity between egfp-nt and nift or blocking enzyme-linked immunosorbent assay (elisa) were both 100%. t ...201323698523
[metagenomics-based detection of swine viruses].extreme varieties of viruses exist in the environment and animals, some of which are unknown. however, many unknown viruses are barely detected by means of conventional virus isolation and pcr assay.201323627112
phylogenetic analysis using e2 gene of classical swine fever virus reveals a new subgenotype in china.outbreaks of classical swine fever (csf) have caused serious economic consequences in china. phylogenetic analysis based on full-length e2 gene sequences showed that five classical swine fever virus (csfv) isolates collected from hunan province in 2011 and 2012, together with seven other isolates from neighboring provinces, guangdong (5) and guangxi (2), could be classified as a new subgenotype 2.1c, which may have been endemic in the south of china for at least fourteen years. subgenotype 2.1c ...201323608662
detection of foot-and-mouth disease serotype o by elisa using a monoclonal antibody.an elisa assay with monoclonal antibody (melisa) was used to type serotype o of foot-and-mouth disease virus (fmdv). all fmdv serotype o reference strains were positive by melisa, while other viruses such as fmdv serotypes asia 1, c, and a and classical swine fever virus, swine vesicular disease virus, and porcine reproductive and respiratory syndrome virus remained negative. furthermore, fmdv serotype o positive samples were able to be detected by melisa. this assay may be particularly suitable ...201323600506
complete genome sequences of classical swine fever virus strains isolated from wild boars in south korea.classical swine fever is a disease that is devastating the pig industry worldwide. here, we report the complete genome sequences of two classical swine virus strains (yc11wb and pc11wb), isolated from korean wild boars in 2011. both strains belong to subgenotype 2.1b. the complete genome sequences of pc11wb and yc11wb are more similar to that of strain zj0801 (isolated in china) than to that of the sw03 strain isolated from domestic pigs in south korea.201323599291
hyper-enhanced production of foreign recombinant protein by fusion with the partial polyhedrin of nucleopolyhedrovirus.to enhance the production efficiency of foreign protein in baculovirus expression systems, the effects of polyhedrin fragments were investigated by fusion expressing them with the enhanced green fluorescent protein (egfp). recombinant viruses were generated to express egfp fused with polyhedrin fragments based on the previously reported minimal region for self-assembly and the krkk nuclear localization signal (nls). fusion expressions with polyhedrin amino acids 19 to 110 and 32 to 110 lead to l ...201323593321
viroporin activity and membrane topology of classic swine fever virus p7 protein.viroporins are a group of viral proteins that participate in viral replication cycles, including modification of membrane permeability and promotion of viral release. although biological data have been accumulated on viroporion-like proteins of other viruses belonging to family flaviviridae, the viroporin activity and membrane topology of p7 protein from classical swine fever virus (csfv), a member of the genus pestivirus of the family flaviviridae, are largely unknown. in this study, sequence a ...201323583663
analysis of classical swine fever virus rna replication determinants using replicons.self-replicating rnas (replicons), with or without reporter gene sequences, derived from the genome of the paderborn strain of classical swine fever virus (csfv) have been produced. the full-length viral cdna, propagated within a bacterial artificial chromosome, was modified by targeted recombination within escherichia coli. rna transcripts were produced in vitro and introduced into cells by electroporation. the translation and replication of the replicon rnas could be followed by the accumulati ...201323580431
[study on using nsp2 protein of porcine reproductive and respiratory syndrome virus (hun4-f112) to express e2 neutralizing epitope of classical swine fever virus].establishment of recombinant porcine reproductive and respiratory syndrome virus (prrsv) with co-expression e2 epitope of classical swine fever virus (csfv) is a crucial step to develop a genetic engineered vaccine against prrsv and csfv. reverse genetic manipulation could be adopted as a com monly used technique. in this study, we focus on using nonessential regions of nsp2 (aa480-532 and aa508-532) as viral vector to express e2 epitope of csfv. a neutralizing epitope of classical swine fever v ...201323547375
multiplex pcr for the simultaneous detection of porcine reproductive and respiratory syndrome virus, classical swine fever virus, and porcine circovirus in pigs.a multiplex polymerase chain reaction (pcr) was designed for the simultaneous detection of three viruses involved in reproductive and respiratory failure in pigs: porcine reproductive and respiratory syndrome virus (prrsv), classical swine fever virus (csfv) and porcine circovirus type 2 (pcv-2). each target produced a specific amplicon with a size of 718 bp (prrsv), 288 bp (csfv), or 466 bp (pcv-2). the sensitivity of the multiplex pcr using purified plasmid constructs containing the specific v ...201323524252
development and partial validation of a recombinant e2-based indirect elisa for detection of specific igm antibody responses against classical swine fever virus.detecting classical swine fever virus specific antibody responses is critical for prevention and control of csf. in this study, a δe2-based indirect elisa was developed to detect specific igm antibodies against csfv. the optimized conditions that were determined experimentally are: a δe2 antigen concentration of 0.5 μg/ml, a serum sample dilution of 1/100 incubated at 37°c for 1.5 h, and a hrp conjugated rabbit anti-pig igm dilution of 1/50,000 incubated at 37°c for 1 h. three hundred clinical s ...201323500647
hemoglobin subunit beta interacts with the capsid protein and antagonizes the growth of classical swine fever virus.the capsid (c) protein of the flaviviridae family members is involved in nucleocapsid formation and virion assembly. however, the influence of c protein-interacting partners on the outcome of pestivirus infections is poorly defined. in this study, hemoglobin subunit beta (hb) was identified as a c protein-binding protein by glutathione s-transferase pulldown and subsequent mass spectrometry analysis of pk-15 cells, which are permissive cells for classical swine fever virus (csfv). coimmunoprecip ...201323487454
conformational and thermal stability improvements for the large-scale production of yeast-derived rabbit hemorrhagic disease virus-like particles as multipurpose vaccine.recombinant virus-like particles (vlp) antigenically similar to rabbit hemorrhagic disease virus (rhdv) were recently expressed at high levels inside pichia pastoris cells. based on the potential of rhdv vlp as platform for diverse vaccination purposes we undertook the design, development and scale-up of a production process. conformational and stability issues were addressed to improve process control and optimization. analyses on the structure, morphology and antigenicity of these multimers we ...201323460801
approaches to define the viral genetic basis of classical swine fever virus virulence.classical swine fever (csf), a highly contagious disease of pigs caused by the classical swine fever virus (csfv), can lead to important economic losses in the pig industry. numerous csfv isolates with various degrees of virulence have been isolated worldwide, ranging from low virulent strains that do not result in any apparent clinical signs to highly virulent strains that cause a severe peracute hemorrhagic fever with nearly 100% mortality. knowledge of the molecular determinants of csfv virul ...201323415391
sequencing approach to analyze the role of quasispecies for classical swine fever.classical swine fever virus (csfv) is a positive-sense rna virus with a high degree of genetic variability among isolates. high diversity is also found in virulence, with strains covering the complete spectrum from avirulent to highly virulent. the underlying genetic determinants are far from being understood. since rna polymerases of rna viruses lack any proof-reading activity, different genome variations called haplotypes, occur during replication. a set of haplotypes is referred to as a viral ...201323415390
complete genome sequences of classical swine fever virus isolates belonging to a new subgenotype, 2.1c, from hunan province, china.two isolates of a new classical swine fever virus (csfv) subgenotype, 2.1c (hnly-2011 and hnsd-2012), were recently isolated from pigs in hunan province, china. the most significant difference in the amino acid sequences of the polyproteins from subgenotypes 2.1a and 2.1b is an spa → tpv amino acid substitution at positions 777 and 779 in the e2 protein.201323405338
cp7_e2alf oral vaccination confers partial protection against early classical swine fever virus challenge and interferes with pathogeny-related cytokine responses.the conventional c-strain vaccine induces early protection against classical swine fever (csf), but infected animals cannot be distinguished from vaccinated animals. the cp7_e2alf marker vaccine, a pestivirus chimera, could be a suitable substitute for c-strain vaccine to control csf outbreaks. in this study, single oral applications of cp7_e2alf and c-strain vaccines were compared for their efficacy to induce protection against a csf virus (csfv) challenge with the moderately virulent bas-rhin ...201323398967
microbiological identification and analysis of swine lungs collected from carcasses in swine farms, china.the primary objective of this 3 years study was to determine the prevalence of porcine pathogens of the lungs of swine in swine farms in southern china. a total of 5,420 samples were collected from 200 swine farms. the bacterium that was most commonly isolated was streptococcus suis, with 10.24 % of the samples being positive, 114 lungs (2.1 %) were positive for pseudorabies virus and 263 (4.85 %) were positive for classical swine fever virus; much lower than positive for prrsv (15.1 %, p = 0.02 ...201324426158
down-regulation of cellular protein heme oxygenase 1 inhibits proliferation of classical swine fever virus in pk-15 cells.heme oxygenase 1 (ho-1) is an inducible enzyme that exerts potent antioxidant and anti-inflammatory effects, which also plays a critical role in host defenses against microbial, and particularly viral, infections. in our previous study, up-regulation of ho-1 was observed in peripheral blood leukocytes (pbls) by genomic expression profiling, following infection of pigs with virulent classical swine fever virus (csfv), the causative agent of a highly contagious disease threatening global pig indus ...201323357296
a serological survey of selected pathogens in wild boar (sus scrofa) in northern turkey.during the hunting season in march 2012, a total of 93 blood samples were collected from wild boars (sus scrofa) shot in the area of northern turkey (samsun and gumushane provinces). these blood samples were examined by enzyme immunoassay (elisa) for the presence of antibodies to classical swine fever virus (csfv), aujeszky's disease virus (adv), porcine reproductive and respiratory syndrome virus (prrsv), porcine respiratory coronavirus (prcv), swine influenza virus (siv), porcine parvovirus (p ...201332214940
pathway analysis in blood cells of pigs infected with classical swine fever virus: comparison of pigs that develop a chronic form of infection or recover.infection of pigs with csfv can lead to either acute disease, resulting in death or recovery, or chronic disease. the mechanisms by which csfv manipulates the pig's first line of defence to establish a chronic infection are poorly understood. therefore, pigs were infected with moderately virulent csfv, and whole blood was collected on a regular basis during a period of 18 days. using whole-genome microarrays, time-dependent changes in gene expression were recorded in blood cells of chronically d ...201323053521
first molecular identification and characterization of classical swine fever virus isolates from nepal.classical swine fever (csf) is a major constraint to pig production worldwide, and in many developing countries, the epidemiological status is unknown. here, for the first time, molecular identification and characterization of csfv isolates from two recent outbreaks in nepal are presented. analysis of full-length e2-encoding sequences revealed that these isolates belonged to csfv subgenotype 2.2 and had highest genetic similarity to isolates from india. hence, for csfv, nepal and india should be ...201322975985
genetic typing of classical swine fever virus isolates from china.the e2 genes of 73 classical swine fever virus (csfv) originated from csf suspected cases in different regions of china were genetically characterized and compared with reference csf viruses. all chinese viruses that characterized were segregated into two major groups and subdivided into four subgroups. most of isolates (61.6%) belonged to group 2 and were further divided into three subgroups: subgroup 2.1, 2.2 and 2.3. subgroup 2.1 was the largest subgroup which contained 46.6% of isolates, whi ...201322672483
baculovirus expression and antigenic characterization of classical swine fever virus e2 proteins.genes encoding a major structural glycoprotein, e2, of classical swine fever viruses (csfv) brescia (subgroup 1.2), paderborn (subgroup 2.1) and kanagawa (subgroup 3.4) were constructed by removing the transmembrane domain and adding a c-terminal 6 histidine (his) tag. all the e2 constructs were efficiently expressed in a baculovirus system as 53-kda glycosylated proteins that were identified in western blots by their reaction with anti-his and csfv-specific antibodies. these proteins were used ...201322510427
classical swine fever virus p7 protein is a viroporin involved in virulence in swine.the nonstructural protein p7 of classical swine fever virus (csfv) is a small hydrophobic polypeptide with an apparent molecular mass of 6 to 7 kda. the protein contains two hydrophobic stretches of amino acids interrupted by a short charged segment that are predicted to form transmembrane helices and a cytosolic loop, respectively. using reverse genetics, partial in-frame deletions of p7 were deleterious for virus growth, demonstrating that csfv p7 function is critical for virus production in c ...201222496228
a novel pcr-based method for high throughput prokaryotic expression of antimicrobial peptide genes.to facilitate the screening of large quantities of new antimicrobial peptides (amps), we describe a cost-effective method for high throughput prokaryotic expression of amps. eddie, an autoproteolytic mutant of the n-terminal autoprotease, npro, from classical swine fever virus, was selected as a fusion protein partner. the expression system was used for high-level expression of six antimicrobial peptides with different sizes: bombinin-like peptide 7, temporin g, hexapeptide, combi-1, human hista ...201222439858
establishment and characterization of an infectious cdna clone of a classical swine fever virus lom strain.classical swine fever virus (csfv) causes a highly contagious disease among swine that has an important economic impact worldwide. csfv strain lom is an attenuated virus of low virulent strain of miyagi isolated from japan in 1956. eight dna fragments representing the genome of the csfv strain lom were obtained by rt-pcr. these were used to determine the complete nucleotide sequence and construct a full-length cdna clone which was called flc-lom. sequence analysis of the recombinant clone (flc-l ...201222437540
characterisation of vaccine-induced, broadly cross-reactive ifn-γ secreting t cell responses that correlate with rapid protection against classical swine fever virus.live attenuated c-strain classical swine fever viruses (csfv) provide a rapid onset of protection, but the lack of a serological test that can differentiate vaccinated from infected animals limits their application in csf outbreaks. since immunity may precede antibody responses, we examined the kinetics and specificity of peripheral blood t cell responses from pigs vaccinated with a c-strain vaccine and challenged after five days with a genotypically divergent csfv isolate. vaccinated animals di ...201222366027
n(pro) of classical swine fever virus prevents type i interferon-mediated priming of conventional dendritic cells for enhanced interferon-α response.a hallmark of acute classical swine fever is the high interferon (ifn)-α levels found in the serum early after infection, followed by an inflammatory cytokine storm. plasmacytoid dendritic cells (pdcs) represent the only known cell type that produces ifn-α upon classical swine fever virus (csfv) infection in vitro. in primary target cells of the virus the viral protein n(pro) inhibits the induction of type i ifn via the degradation of irf3. we hypothesized that the early systemic pdc-derived ifn ...201222313263
yeast expressed classical swine fever e2 subunit vaccine candidate provides complete protection against lethal challenge infection and prevents horizontal virus transmission.classical swine fever (csf) caused by the classical swine fever virus (csfv) is a highly contagious swine disease resulting in large economical losses worldwide. the viral envelope glycoprotein e(rns) and e2 are major targets for eliciting antibodies against csfv in infected animals. a pichia pastoris yeast expressed e2 protein (ye2) has been shown to induce a protective immune response against csfv challenge. the purpose of this study is to determine the optimal dose of ye2 and its efficacy on ...201222300723
classical swine fever virus ns5a protein interacts with 3'-untranslated region and regulates viral rna synthesis.to investigate the function of classical swine fever virus (csfv) ns5a protein, the experiments for viral rna synthesis and viral replication were performed in the co-presence of ns5a and ns5b. results showed that small concentrations of ns5a stimulated, large concentrations of ns5a inhibited, viral rna synthesis and viral replication. affinity chromatography experiments and uv-crosslinking assays revealed that csfv ns5a and ns5b bound its cognate 3'utr and that ns5a had higher affinity than ns5 ...201222261205
effects of astragalus polysaccharide on immune responses of porcine pbmc stimulated with prrsv or csfv.astragalus polysaccharide (aps) has been used as an immunomodulator that can enhance immune responses, whereas the immunomodulatory effects of aps on porcine peripheral blood mononuclear cells (pbmcs) exposed to porcine reproductive and respiratory syndrome virus (prrsv) and classical swine fever virus (csfv) have not been investigated.201222253710
[quantitative detection of anti-classical swine fever virus sirna expression in cells by stem-loop rt-qpcr].rna interference (rnai) is a promising technology in development of specific antiviral therapy, but the quantitative detection of small interfering rna (sirna) expressed in vivo is the main challenge to assess its antiviral effect. in order to detect the sirna molecules (sin1 and sin2) particularly expressed in cells to inhibit the replication of classical swine fever virus (csfv), serial specific stem-loop primers were designed and synthesized. two of them (slp-n1-6 and slp-n2-8) were selected ...201222667106
real-time analysis of the interaction of a multiple-epitope peptide with antibodies against classical swine fever virus using surface plasmon resonance.the e2 envelope glycoprotein is the major immunodominant protein of classical swine fever virus (csfv), and can induce neutralizing antibodies and protective immune responses in infected swine. we developed a tandem-repeat multiple-epitope recombinant protein that contains two copies of each of the regions of e2 spanned by residues 693-704, 770-780, and 826-843, coupled by two copies of the region spanned by residues 1446-1460 of the csfv nonstructural protein ns2-3. the chemically synthesized g ...201222634031
withdrawn: development of an rt-pcr-rflp assay for the detection and differentiation of wild-type and vaccine strains of classical swine fever virus.this article has been withdrawn at the request of the author(s). the publisher apologizes for any inconvenience this may cause. the full elsevier policy on article withdrawal can be found at http://www.elsevier.com/locate/withdrawalpolicy.201222609180
early pathogenesis of classical swine fever virus (csfv) strains in danish pigs.host-virus interactions play an important role for the clinical outcome of classical swine fever virus (csfv) infections in pigs. strain virulence, host characteristics and environment are all factors that markedly influence disease severity. we tested csfv strains of varying virulence in an experimental set-up, reducing the influence of host and environmental factors. thus, weaner pigs were inoculated with one of 4 csfv strains in order to compare the pathogenesis for a 3-week-period after infe ...201222608103
positive selection pressure on the b/c domains of the e2-gene of classical swine fever virus in endemic areas under c-strain vaccination.in cuba, classical swine fever (csf) has become an endemic disease with several outbreaks each year, despite the implemented vaccination program. interestingly, a trend towards a milder presentation of the disease has been observed among the animals during the last years. this study aimed to assess positive selection pressure acting on partial e2 gene of csf viruses to gain insights into the mechanisms governing virulence and the driving forces of classical swine fever virus (csfv) evolution in ...201222580241
development of multiplex pcr for simultaneous detection of six swine dna and rna viruses.uniplex and multiplex reverse transcription-polymerase chain reaction (rt-pcr) and pcr protocols were developed and evaluated subsequently for its effectiveness in detecting simultaneously single and mixed infections in swine. specific primers for three dna viruses and three rna viruses, including classical swine fever virus (csfv), porcine reproductive and respiratory syndrome virus (prrsv), japanese encephalitis virus (jev), porcine circovirus type 2 (pcv2), porcine pseudorabies virus (prv) an ...201222575688
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