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the african swine fever virus dp71l protein recruits the protein phosphatase 1 catalytic subunit to dephosphorylate eif2alpha and inhibits chop induction but is dispensable for these activities during virus infection.the african swine fever virus (asfv) dp71l protein is present in all isolates as either a short form of 70 to 72 amino acids or a long form of about 184 amino acids, and both of these share sequence similarity to the c-terminal domain of the herpes simplex virus icp34.5 protein and cellular protein gadd34. in the present study we expressed dp71l in different mammalian cells and demonstrated that dp71l causes dephosphorylation of eukaryotic translation initiation factor 2 alpha (eif2α) in resting ...201020702639
matrix-assisted refolding of autoprotease fusion proteins on an ion exchange column: a kinetic investigation.matrix-assisted refolding is an excellent technique for performing refolding of recombinant proteins at high concentration because aggregation during refolding is partially suppressed. the autoprotease n(pro) and its engineered mutant eddie can be efficiently refolded on cation-exchangers. in the current work, denatured fusion proteins were loaded at different column saturations (5 and 50 mg ml(-1) gel), and refolding and self-cleavage were initiated during elution. the contact time of the prote ...201020708192
[construction and immunogenicity of a recombinant adenovirus co-expressing the e2 protein of classical swine fever virus and the porcine interleukin 2 in rabbits].to construct a recombinant adenovirus co-expressing the e2 protein of classical swine fever virus (csfv) and the porcine interleukin 2 (pil-2), the csfv e2 gene and pil-2 gene were amplified respectively from the plasmids pmd19-t-e2 and pmd19-t-pil-2 by pcr. e2-pil-2 fusion gene was obtained by using 5 consecutive glycine codons as a linker and cloned into the adenoviral shuttle plasmid adtrack. the adtrack-e2-pil-2 was linearized and transformed into e. coli bj5183 with the backbone plasmid ade ...201021043139
development of a loop-mediated isothermal amplification for visual detection of the hclv vaccine against classical swine fever in china.a loop-mediated isothermal amplification (lamp) assay was developed and evaluated for the rapid and specific detection of hclv vaccine strain against classical swine fever. four primers were designed for amplification of ns5b gene region with bst dna polymerase at a constant temperature of 65°c. the products showed ladder-like pattern on 2% agarose gel, and can be visualised after addition of sybr green i dye. the detection limit of the assay was 5 copies of the hclv genome per reaction. no cros ...201021055419
the african swine fever virus lectin ep153r modulates the surface membrane expression of mhc class i antigens.we have modeled a 3d structure for the c-type lectin domain of the african swine fever virus protein ep153r, based on the structure of cd69, cd94 and ly49a cell receptors, and this model predicts that a dimer of ep153r may establish an asymmetric interaction with one mhc-i molecule. a functional consequence of this interaction could be the modulation of mhc-i expression. by using both transfection and virus infection experiments, we demonstrate here that ep153r inhibits mhc-i membrane expression ...201021069396
structural models of dynll1 with interacting partners: african swine fever virus protein p54 and postsynaptic scaffolding protein gephyrin.dynll1, the smallest dynein light chain, interacts with different cargos facilitating their cellular transport. usually the sequence recognized in the targets is homologous to the giqvd or the kxtqt motifs with a glutamine that is important for binding. here we add two new examples of dynll1 targets that can be classified into these two groups: asfv p54 and gephyrin. using nmr we demonstrate the direct interaction between dynll1 and two peptides derived from their interacting sequences. we model ...201021094642
development of a highly sensitive real-time rt-pcr protocol for the detection of classical swine fever virus independent of the 5' untranslated region.classical swine fever (csf) is one of the most severe diseases of pigs, and can cause immense economic losses. real-time reverse transcription polymerase chain reaction (rrt-pcr) can be used as a sensitive and specific method for detection of classical swine fever virus (csfv). different published protocols are used routinely for csfv diagnosis. however, almost all these systems use the highly conserved 5' untranslated region (5'utr) of the csfv genome as template. for a reliable diagnosis in ou ...201021111760
first detection of african swine fever virus in ornithodoros porcinus in madagascar and new insights into tick distribution and taxonomy.abstract:201021118485
emergence of african swine fever virus, northwestern iran.in 2008, african swine fever was introduced into georgia, after which it spread to neighboring armenia, azerbaijan, and the russian federation. that same year, pcr and sequence analysis identified african swine fever virus in samples from 3 dead female wild boars in northwestern iran. wild boars may serve as a reservoir.201021122227
detection of porcine parvovirus using a taqman-based real-time pcr with primers and probe designed for the ns1 gene.a taqman-based real-time polymerase chain reaction (pcr) assay was devised for the detection of porcine parvovirus (ppv). two primers and a taqman probe for the non-structural protein ns1 gene were designed. the detection limit was 1 x 10² dna copies/μl, and the assay was linear in the range of 1 x 10² to 1 x 10⁹ copies/μl. there was no cross-reaction with porcine circovirus 2 (pcv2), porcine reproductive and respiratory syndrome virus (prrsv), pseudorabies virus (prv), classical swine fever vir ...201021126330
time-dependent infection probability of classical swine fever via excretions and secretions.several routes contribute to the spread of classical swine fever (csf) during outbreaks of this disease. however, for many infected herds in recent epidemics, no route of virus introduction could be indentified. to obtain more insight into the relative importance of secretions and excretions in transmission of csf virus, a model was developed. this model quantified the daily transmission probabilities from one infectious pig to one susceptible pig, using quantitative data on: (a) virus excretion ...201021145604
novel bioluminescent quantitative detection of nucleic acid amplification in real-time.the real-time monitoring of polynucleotide amplification is at the core of most molecular assays. this conventionally relies on fluorescent detection of the amplicon produced, requiring complex and costly hardware, often restricting it to specialised laboratories.201021152399
design and verification of a highly reliable linear-after-the-exponential pcr (late-pcr) assay for the detection of african swine fever virus.african swine fever virus (asfv) is a highly pathogenic dna virus that is the causative agent of african swine fever (asf), an infectious disease of domestic and wild pigs of all breeds and ages, causing a range of syndromes. acute disease is characterized by high fever, haemorrhages in the reticuloendothelial system, and a high mortality rate. a powerful novel diagnostic assay based on the linear-after-the-exponential-pcr (late-pcr) principle was developed to detect asfv. late-pcr is an advance ...201021167207
experimental infection of common warthogs (phacochoerus africanus) and bushpigs (potamochoerus larvatus) with classical swine fever virus ii: a comparative histopathological study.wild african suidae, the common warthog (phacochoerus africanus) and bushpig (potamochoerus larvatus), were experimentally infected with classical swine fever (csf) virus following the diagnosis of csf subtype 2.1 in domestic pigs in south africa in 2005. no data regarding the susceptibility or potential lesions of these african wild suids are available. seven subadult warthogs and six bushpigs were captured and infected intranasally with the south african isolate. two in-contact control animals ...201021176120
phenotypic and cytologic studies of lymphoid cells and monocytes in primary culture of porcine bone marrow during infection of african swine fever virus.we have modeled in vitro infection of african swine fever virus (asfv) in primary unstimulated cells of the porcine bone marrow and have studied the phenotypical changes in the population of porcine lymphoid cells by cytophotometry. monocytes and large-sized lymphocytes completely vanished in 72 h of infection which is result of high sensitivity of those cells to asfv. we describe dna synthesis in monocytes at 24 h post infection. cytophotometry of the uninfected cells revealed the few number of ...201021184199
immunogenic and replicative properties of classical swine fever virus replicon particles modified to induce ifn-α/β and carry foreign genes.virus replicon particles (vrp) are genetically engineered infectious virions incapable of generating progeny virus due to partial or complete deletion of at least one structural gene. vrp fulfil the criteria of a safe vaccine and gene delivery system. with vrp derived from classical swine fever virus (csf-vrp), a single intradermal vaccination protects from disease. spreading of the challenge virus in the host is however not completely abolished. parameters that are critical for immunogenicity o ...201021184857
a multiplex rt-pcr for rapid and simultaneous detection of porcine teschovirus, classical swine fever virus, and porcine reproductive and respiratory syndrome virus in clinical specimens.a multiplex rt-pcr (mrt-pcr) assay was developed and evaluated for its ability to detect multiple viral infections of swine simultaneously. one pair of primers was selected carefully for each of the following three rna viruses: porcine reproductive and respiratory syndrome virus (prrsv), classical swine fever virus (csfv), and porcine teschovirus (ptv). each target produced a specific amplicon with a size of 451bp (prrsv), 343bp (csfv), or 163bp (ptv). the sensitivity of the mrt-pcr using purifi ...201021192983
antigenic analysis of classical swine fever virus e2 glycoprotein using pig antibodies identifies residues contributing to antigenic variation of the vaccine c-strain and group 2 strains circulating in china.glycoprotein e2, the immunodominant protein of classical swine fever virus (csfv), can induce neutralizing antibodies and confer protective immunity in pigs. our previous phylogenetic analysis showed that subgroup 2.1 viruses branched away from subgroup 1.1, the vaccine c-strain lineage, and became dominant in china. the e2 glycoproteins of csfv c-strain and recent subgroup 2.1 field isolates are genetically different. however, it has not been clearly demonstrated how this diversity affects anti ...201021194462
proteomic analysis of primary porcine endothelial cells after infection by classical swine fever virus.endothelial cells are the main target of classical swine fever virus during infection, and extensive hemorrhage is the most typical clinical sign of classical swine fever. to investigate the molecular mechanism of hemorrhagic pathogenesis, two-dimensional difference gel electrophoresis with fluorescent dyes (2d-dige) was used to analyze the proteomic profile of primary porcine umbilical vein endothelial cells (puvecs) following csfv infection. of 15 protein spots with differential expression, 8 ...201020595071
sensitive detection of african swine fever virus using real-time pcr with a 5' conjugated minor groove binder probe.the design of a 5' conjugated minor groove binder (mgb) probe real-time pcr assay is described for the rapid, sensitive and specific detection of african swine fever virus (asfv) dna. the assay is designed against the 9gl region and is capable of detecting 20 copies of a dna standard. it does not detect any of the other common swine dna viruses tested in this study. the assay can detect asfv dna in a range of clinical samples. sensitivity was equivalent to the office international des epizooties ...201020621646
expression and immunological studies of classical swine fever virus glycoprotein e2 in the bi-cistronic baculovirus/larvae expression system.to develop an economical, easy technique for producing recombinant e2 glycoprotein (re2) of classical swine fever virus (csfv) as a candidate immunogen, a bi-cistronic baculovirus/larvae expression vector was constructed using p10 promoter, an internal ribosome entry site, and the gfp gene. trichoplusia ni larvae were successfully infected with the occluded recombinant baculovirus via feed, and the characteristics of re2 were confirmed by immunoblot and glycosylation stain. re2 at a concentratio ...201020622456
the classical swine fever virus n-terminal protease n(pro) binds to cellular hax-1.the positive-stranded rna genome of classical swine fever virus (csfv) encodes 12 known proteins. the first protein to be translated is the n-terminal protease (n(pro)). n(pro) helps evade the innate interferon response by targeting interferon regulatory factor-3 for proteasomal degradation and also participates in the evasion of dsrna-induced apoptosis. to elucidate the mechanisms by which n(pro) functions, we performed a yeast two-hybrid screen in which the anti-apoptotic protein hax-1 was ide ...201020631090
classical swine fever virus ns3 is an ires-binding protein and increases ires-dependent translation.to get more evidences for understanding the role of ns3 in viral translation, we observed the promotive effect of csfv ns3 on ires-mediated translation by using dicistronic and monocistronic systems containing the precise segment comprising csfv ires. the results for affinity chromatography and uv-crosslinking assays indicated that ns3 bound csfv ires and that csfv ns5a and ns5b could reduce the ires-ns3 interaction. further experiments showed that the ns5a also bound the ires and that ns3 and n ...201020637813
complete genomic sequence of a border disease virus isolated from pyrenean chamois.this report describes the full-length genome sequence of the pestivirus strain h2121 which was recently isolated from pyrenean chamois and typed as border disease virus (bdv) genotype 4. comparison with full-length genomic sequences of the approved pestivirus species bovine viral diarrhea virus-1 (bvdv-1), bvdv-2, bdv, and classical swine fever virus, the tentative species represented by strain giraffe-1, as well as the atypical pestivirus strain th/04_khonkaen confirmed that the chamois pestivi ...201020638945
molecular epidemiology of current classical swine fever virus isolates of wild boar in germany.classical swine fever (csf) has caused significant economic losses in industrialized pig production, and is still present in some european countries. recent csf outbreaks in europe were mainly associated with strains of genogroup 2 (subgroup 2.3). although there are extensive datasets regarding 2.3 strains, there is very little information available on longer fragments or whole classical swine fever virus (csfv) genomes. furthermore, there are no detailed analyses of the molecular epidemiology o ...201020660149
classical swine fever virus in south-eastern europe--retrospective analysis of the disease situation and molecular epidemiology.classical swine fever (csf) is among the most important diseases of domestic pigs and causes great socio-economic losses. therefore, control of csf is given high priority within the european union, including financial support of concerted control actions in candidate and in potential candidate countries. unfortunately, from some of these countries information on the csf situation and related data is very limited. this study was undertaken to gather all available information on the domestic pig p ...201020541876
crystal structure and functional characterization of an immunomodulatory salivary cystatin from the soft tick ornithodoros moubata.the saliva of blood-feeding parasites is a rich source of peptidase inhibitors that help to overcome the host's defence during host-parasite interactions. using proteomic analysis, the cystatin omc2 was demonstrated in the saliva of the soft tick ornithodoros moubata, an important disease vector transmitting african swine fever virus and the spirochaete borrelia duttoni. a structural, biochemical and biological characterization of this peptidase inhibitor was undertaken in the present study. rec ...201020545626
phylogenetic analysis of classical swine fever virus isolates from peru.classical swine fever (csf) is considered to be endemic in peru with outbreaks reported to the world organization for animal health as recently as 2008 and 2009. nevertheless, little is known regarding the genetic subgroup(s) of csf virus that are circulating in peru or their relationship to recent csf viruses that have been isolated from neighbouring south american countries or other parts of the world. in this study, we molecularly characterize csf viruses that were isolated from domestic pigs ...201020545910
understanding inhibition of viral proteins on type i ifn signaling pathways with modeling and optimization.the interferon system provides a powerful and universal intracellular defense mechanism against viruses. as one part of their survival strategies, many viruses have evolved mechanisms to counteract the host type i interferon (ifn-alpha/beta) responses. in this study, we attempt to investigate virus- and double-strand rna (dsrna)-triggered type i ifn signaling pathways and understand the inhibition of ifn-alpha/beta induction by viral proteins using mathematical modeling and quantitative analysis ...201020553733
identification of antigen-specific residues on e2 glycoprotein of classical swine fever virus.envelope glycoprotein e2 of classical swine fever virus (csfv) is the major antigen that induces neutralizing antibodies in infected pigs. our previous study revealed that n-terminal 90 residues (domains b/c) of e2 play key roles in differentiating vaccine strain lpc/ahri (subgroup 1.1) from the two field strains td/96/twn (subgroup 2.1) and 94.4/il/94/twn (subgroup 3.4) (chang et al., 2010). this study further analyzed the reaction patterns between monoclonal antibodies (mabs) and expressed hyb ...201020558217
dynamin- and clathrin-dependent endocytosis in african swine fever virus entry.african swine fever virus (asfv) is a large dna virus that enters host cells after receptor-mediated endocytosis and depends on acidic cellular compartments for productive infection. the exact cellular mechanism, however, is largely unknown. in order to dissect asfv entry, we have analyzed the major endocytic routes using specific inhibitors and dominant negative mutants and analyzed the consequences for asfv entry into host cells. our results indicate that asfv entry into host cells takes place ...201019939916
a one-step real-time reverse transcription-polymerase chain reaction detection of classical swine fever virus using a minor groove binding probe.the aim of this study was to develop a one-step real-time reverse transcription-polymerase chain reaction assay using the minor groove binding probe (mgb rrt-pcr) for rapid and quantitative detection of classical swine fever virus (csfv). the method, which targets the 5'-nontranslated region (5'ntr) of the viral genome, detected all csfv isolate tested, but not heterologous pathogens. using an in vitro transcript of the 5'ntr as a quantitative standard for the csfv genome copy number, the assay ...201020411415
intranuclear detection of african swine fever virus dna in several cell types from formalin-fixed and paraffin-embedded tissues using a new in situ hybridisation protocol.in this study, a new in situ hybridisation (ish) protocol has been developed to identify african swine fever virus (asfv) genome in formalin-fixed, paraffin-embedded tissues. different digoxigenin labelled asfv-probes were tested, including single asfv-specific oligonucleotides, an 18.5kb restriction fragment from the viral genome and the entire asfv genome. the latter showed the highest sensitivity in all tissues tested, independently of the virus used for challenge: e75l or ba71l. although a s ...201020417663
enhanced immunity against classical swine fever in pigs induced by prime-boost immunization using an alphavirus replicon-vectored dna vaccine and a recombinant adenovirus.classical swine fever (csf) - caused by the classical swine fever virus (csfv) - is a fatal disease of pigs that is responsible for extensive losses to the swine industry worldwide. we had demonstrated previously that a prime-boost vaccination strategy using an alphavirus (semliki forest virus, sfv) replicon-vectored dna vaccine (psfv1cs-e2) and a recombinant adenovirus (radv-e2) expressing the e2 glycoprotein of csfv induced enhanced immune responses in a mouse model. in this study, we evaluate ...201020435352
a study on the applicability of implantable microchip transponders for body temperature measurements in pigs.the applicability of an electronic monitoring system using microchip transponders for measurement of body temperatures was tested in 6-week-old conventional danish weaners infected with classical swine fever virus (csfv). subcutaneous tissue temperatures obtained by the implantable transponders were compared with rectal temperatures, recorded by a conventional digital thermometer.201020444254
the development of a rapid sybr one step real-time rt-pcr for detection of porcine reproductive and respiratory syndrome virus.prompt detection of prrsv in the field samples is important for effective prrs control, thereby reducing the potentially serious economic damage which can result from an outbreak. in this study, a rapid sybr-based, one step real-time rt-pcr quantitative reverse transcription pcr (qrt-pcr) has been developed for the detection of porcine reproductive and respiratory syndrome virus (prrsv). primers were designed based on the sequence of highly conservative region of prrsv n gene.201020459705
the n-glycosylation of classical swine fever virus e2 glycoprotein extracellular domain expressed in the milk of goat.classical swine fever virus (csfv) outer surface e2 glycoprotein represents an important target to induce protective immunization during infection but the influence of n-glycosylation pattern in antigenicity is yet unclear. in the present work, the n-glycosylation of the e2-csfv extracellular domain expressed in goat milk was determined. enzymatic n-glycans releasing, 2-aminobenzamide (2ab) labeling, weak anion-exchange and normal-phase hplc combined with exoglycosidase digestions and mass spect ...201020460099
optimisation of reverse transcription can improve the sensitivity of rt-pcr for detection of classical swine fever virus.classical swine fever is a highly contagious, notifiable disease of pigs and wild boars listed by the world organisation for animal health (oie). therefore, methods employed in the diagnosis of csf should be fast, sensitive and specific. the aim of this study was optimisation of the reverse transcription reaction to increase the sensitivity of real-time rt-pcr for the detection of classical swine fever virus, the aetiological agent of the disease. the efficiency of reverse transcription reaction ...201020460224
changes in the porcine peripheral blood mononuclear cell proteome induced by infection with highly virulent classical swine fever virus.leukopenia and immunosuppression are characteristic clinical manifestations of classical swine fever and peripheral blood mononuclear cells (pbmcs) are major targets of classical swine fever virus. to investigate proteomic expression changes in swine pbmcs during lethal csfv infection, proteins of pbmcs from five lethally csfv-infected pigs were resolved by two-dimensional electrophoresis followed by mass spectrometry. quantitative intensity analysis revealed that 66 protein spots showed altered ...201020463149
the toll-like receptor adaptor molecule trif enhances dna vaccination against classical swine fever.to evaluate the effects of the toll/interleukin-1 receptor domain-containing adaptor-inducing interferon-beta (trif) on immune responses induced by dna vaccines, mice were immunized with the eukaryotic expression plasmid pcdna/e2 encoding classical swine fever virus (csfv) e2 alone or in combination with the trif genetic adjuvant. immune responses were examined in immunized mice. our data demonstrates that co-delivery of the dna vaccine pcdna/e2 with the trif adjuvant augmented specific humoral ...201020466439
evaluation of a primer-probe energy transfer real-time pcr assay for detection of classical swine fever virus.this study describes evaluation of a real-time pcr assay based on primer-probe energy transfer (priproet) technology for detection of classical swine fever virus (csfv). the priproet technology allows melting curve analysis following pcr amplification and thus provides a higher specificity. the assay was compared with a taqman assay by testing a total of 203 samples including 175 clinical specimens and 28 batches of hog cholera lapinized virus (hclv) vaccine. the two assays gave the same results ...201020471428
classical swine fever virus down-regulates endothelial connexin 43 gap junctions.classical swine fever is a contagious disease of pigs characterized by fatal hemorrhagic fever. classical swine fever virus (csfv) induces the expression of pro-inflammatory and pro-coagulant factors of vascular endothelial cells and establishes a long-term infection. this study aimed to understand the effect of csfv on endothelial connexin 43 (cx43) expression and gap junctional intercellular coupling (gjic). porcine aortic endothelial cells were infected with csfv at different multiplicity of ...201020473696
african swine fever virus protein p17 is essential for the progression of viral membrane precursors toward icosahedral intermediates.the first morphological evidence of african swine fever virus (asfv) assembly is the appearance of precursor viral membranes, thought to derive from the endoplasmic reticulum, within the assembly sites. we have shown previously that protein p54, a viral structural integral membrane protein, is essential for the generation of the viral precursor membranes. in this report, we study the role of protein p17, an abundant transmembrane protein localized at the viral internal envelope, in these process ...201020504920
expression and purification of antimicrobial peptide cm4 by npro fusion technology in e. coli.antimicrobial peptide cm4 is a small cationic peptide with broad-spectrum activities against bacteria, fungi, and tumor cells. different strategies have been developed to produce small antibacterial peptides using recombinant techniques. to date, no efforts to obtain large quantities of active recombinant cm4 have been reported. in order to establish a bacterium-based cm4 production system, cm4 was cloned into pet28a and expressed with npro mutant (eddie) fusion. cm4 expressed as eddie are depos ...201020512388
[progress in dna vaccines against classical swine fever: a review].in 1990, it was reported that the naked dna encoding an antigen (so-called dna vaccine) transduced directly into the muscle is able to induce immune responses just like antigen inoculation. since then, a number of dna vaccines against different diseases have been developed and shown to induce different levels of specific humoral and/or cell-mediated immunity. efforts have been made to develop effective dna vaccines against classical swine fever (csf). this review covered the following aspects in ...201020518338
[sequence analysis, expression and antigenicity detection of bovine viral diarrhea virus ns3 gene].in this study, we cloned the ns3 gene from bovine viral diarrhea virus (bvdv) vedevac strain. the result showed that the average p-distance of pestivirus ns3 amino acid sequence was 0.07 and the vedevac strain was classified to bvdv type 1. using pet-30a(+) as vector and escherichia coli rosetta (de3) as host, we obtained purified recombinant ns3 protein by ni-nta affinity chromatography. western blotting analysis demonstrated that both bvdv positive serum and classical swine fever virus (csfv) ...201020518342
interferon-gamma induction correlates with protection by dna vaccine expressing e2 glycoprotein against classical swine fever virus infection in domestic pigs.classical swine fever (csf) is a highly contagious viral infection affecting domestic and wild pigs. for classical swine fever virus (csfv), immunization with plasmids expressing different versions of glycoprotein e2 has proven an effective way to induce protection. previously, we have also shown that immunization with dna vaccine expressing glycoprotein e2 (dna-e2) induced specific t helper cell responses in the absence of neutralizing antibodies. however, the role of t cell responses in protec ...201019896784
the classical swine fever virus npro product is degraded by cellular proteasomes in a manner that does not require interaction with interferon regulatory factor 3.classical swine fever is a notifiable disease of pigs. the causative agent, classical swine fever virus (csfv), is highly contagious and causes mild to severe haemorrhagic disease depending on the virulence of the strain. the rna genome of csfv is translated as a single polyprotein that is processed to yield 12 proteins. like other pestiviruses, the first protein to be translated is the n-terminal autoprotease termed n(pro). a novel pestiviral protein with no known cellular homologues, n(pro) an ...201019906943
development and validation of reverse transcription loop-mediated isothermal amplification for detection of prrsv.to establish a reverse-transcription loop-mediated isothermal amplification (rt-lamp) method for rapid detection of porcine reproductive and respiratory syndrome virus (prrsv), four primers specific to six regions of the n gene were designed. after amplification in an isothermal water bath for 1 h, samples containing prrsv generated the expected ladder-like products while porcine parvovirus, porcine circovirus, classic swine fever virus, pseudorabies virus, and swine testis cells generated no pr ...201019911264
mutations in classical swine fever virus ns4b affect virulence in swine.ns4b is one of the nonstructural proteins of classical swine fever virus (csfv), the etiological agent of a severe, highly lethal disease of swine. protein domain analysis of the predicted amino acid sequence of the ns4b protein of highly pathogenic csfv strain brescia (bicv) identified a putative toll/interleukin-1 receptor (tir)-like domain. this tir-like motif harbors two conserved domains, box 1 and box 2, also observed in other members of the tir superfamily, including toll-like receptors ( ...201019923180
development and evaluation of rapid detection of classical swine fever virus by reverse transcription loop-mediated isothermal amplification (rt-lamp).a one-step reverse transcription loop-mediated isothermal amplification (rt-lamp) was developed for the detection of classical swine fever virus (csfv) and a set of primers designed based on the e2 gene reference sequences of the csfv. the assay was optimized to amplify csfv rna by incubation at 63 degrees c for 50 min. the rt-lamp amplification products had a ladder-like appearance when electrophoresed on an agarose gel. the rt-lamp assay showed higher sensitivity than the conventional rt-pcr, ...201019931578
characterisation of experimental infections of domestic pigs with genotype 2.1 and 3.3 isolates of classical swine fever virus.the early identification of classical swine fever epizootics is hampered by difficulties in recognising early signs of infection, due to a lack of specific clinical signs. in addition many textbook descriptions of csf are based on observations of disease caused by historic, mainly genotype 1, strains. our objective was to improve our knowledge of the diverse range of signs that different csfv strains can cause by characterising the experimental infection of domestic pigs with both a recent strai ...201019875252
bovine viral diarrhea virus infections in heterologous species.infections with bovine viral diarrhea virus (bvdv) are not limited to cattle, but may be detected in various species in the mammalian order artiodactyla. despite epidemiological evidence of bvdv infections in species other than cattle, current knowledge regarding the impact of bvdv on heterologous species is incomplete. in heterologous hosts, bvdv infections with clinical signs analogous to those in cattle have been described and include disease of multiple organ systems, most notably the reprod ...201019883524
characterization of a new chimeric marker vaccine candidate with a mutated antigenic e2-epitope.a new chimeric pestivirus "cp7_e1e2alf_tla", based on the infectious cdna of bovine viral diarrhea virus (bvdv) strain cp7, was constructed. the substitution of bvdv e1 and e2 with the respective proteins of classical swine fever (csf) strain alfort 187 allows an optimal heterodimerization of e1 and e2 in the chimeric virus, which is beneficial for efficient and authentic virus assembly and growth. in addition, for implementation of e2-based marker diagnostics, the previously described antigenic ...201019892497
classical swine fever virus ns3 enhances rna-dependent rna polymerase activity by binding to ns5b.ns3 of pestiviruses contains a protease domain and a rna helicase/ntpase domain. contradictory results have been reported regarding ns3 in rna synthesis. to investigate the effect of ns3 on classical swine fever virus (csfv) ns5b rna-dependent rna polymerase activity (rdrp) activity and ns3-ns5b interaction, rdrp reactions, gst-pull-down assays and co-immunoprecipitation analyses containing ns5b and either of ns3 protein and the different truncated ns3 mutants were performed, respectively. we fo ...201019951725
the use of cos-1 cells for studies of field and laboratory african swine fever virus samples.different naturally occurring, cell adapted or genetically manipulated stocks of african swine fever virus were able to infect directly cultures of cos-1 cells, producing extensive cytopathic effects and amounts from 10(6) to 10(7) of infective progeny virus per ml. the induction of late virus-specific proteins, demonstrated by rt-pcr and immunoblotting, and the development of lysis plaques by all the virus samples tested so far, allowed the optimization of both titration and diagnostic assays, ...201019961878
detection of african swine fever virus by loop-mediated isothermal amplification.a loop-mediated isothermal amplification (lamp) assay was developed for the detection of african swine fever virus (asfv). this assay targets the topoisomerase ii gene of asfv and its specificity was confirmed by restriction enzyme digestion of the reaction products. the analytical sensitivity of this asfv lamp assay was at least 330 genome copies, and the test was able to detect representative isolates of asfv (n=38) without cross-reacting with classical swine fever virus. the performance of th ...201019963011
classical swine fever virus infection protects aortic endothelial cells from pipc-mediated apoptosis.classical swine fever virus (csfv) causes severe disease in pigs associated with leukopenia, haemorrhage and fever. we show that csfv infection protects endothelial cells from apoptosis induced by the dsrna mimic, pipc, but not from other apoptotic stimuli, fasl or staurosporine. csfv infection inhibits pipc-induced caspase activation, mitochondrial membrane potential loss and cytochrome c release as well as the pro-apoptotic effects of truncated bid (tbid) overexpression. the csfv proteins n(pr ...201020007358
first report of porcine circovirus type 2 infections in cuba.to obtain information about the porcine circovirus type 2 (pcv2) infection status of pigs in cuba and the probable association of pcv2 with other porcine viruses, tissue samples collected from ill pigs were evaluated using polymerase chain reaction (pcr). the pcr analysis showed that 67.7% of the samples (23/34) from seven swine herds of six different geographic regions were detected to be positive for pcv2. ten of the 23 pcv2 positive samples (43.5%) shown a concurrent infection with porcine pa ...201020031180
global transcriptional profiles in peripheral blood mononuclear cell during classical swine fever virus infection.classical swine fever virus (csfv) is an etiologic agent that causes a highly contagious disease in pigs. laying a foundation to solve problems in its pathogenic mechanism, microarray analysis was performed to detect the gene transcriptional profiles in peripheral blood mononuclear cells (pbmc) following infection with a chinese highly virulent csfv strain shimen. three susceptible pigs were inoculated intramuscularly with a lethal dose (1.0x10(6) tcid(50)) of csfv. pigs showed classical csf sig ...201020034523
correlation of the virulence of csfv with evolutionary patterns of e2 glycoprotein.infection with classical swine fever virus (csfv) is costly to the livestock industry. several genomic sequences including velogenic strains and low virulent strains have been identified. however, the reasons for the virulence of the virus have remained unclear. based on selective pattern and pressure strength, we classified all genes of csfv into three classes. among these genes, the e2 gene was under the strongest positive selection. based on the analysis of 85 representative e2 gene sequences ...201020036871
classic swine fever virus ns2 protein leads to the induction of cell cycle arrest at s-phase and endoplasmic reticulum stress.classical swine fever (csf) caused by virulent strains of classical swine fever virus (csfv) is a haemorrhagic disease of pigs, characterized by disseminated intravascular coagulation, thrombocytopoenia and immunosuppression, and the swine endothelial vascular cell is one of the csfv target cells. in this report, we investigated the previously unknown subcellular localization and function of csfv ns2 protein by examining its effects on cell growth and cell cycle progression.201020064240
prevalence of antibodies to selected viral and bacterial pathogens in wild boar (sus scrofa) in campania region, italy.serum samples were collected from wild boars (sus scrofa) harvested during the 2005-2006 hunting season in campania, southern italy. samples were tested for antibodies to leptospira interrogan, brucella spp., salmonella spp., aujeszky disease virus (adv), porcine reproductive and respiratory stress syndrome virus (prrsv), porcine parvovirus (ppv), classical swine fever virus (csfv), and swine vesicular disease virus (svdv). of the 342 serum samples tested, 15 (4.4%) were seropositive to brucella ...201020090052
antigenic domains analysis of classical swine fever virus e2 glycoprotein by mutagenesis and conformation-dependent monoclonal antibodies.glycoprotein e2 of classical swine fever virus (csfv) is the major antigenic protein exposed on the outer surface of the virion that induces main neutralizing antibodies during infection in pigs. this study displays the differences in antigenicity of e2 between vaccine and field strains of csfv by their variable reaction patterns between expressed proteins and monoclonal antibodies (mabs). the d/a domains of various csfvs were relatively conserved and recognized by all mabs against the a domain. ...201020132851
in vitro antiviral activity of some uridine derivatives of 2-deoxy sugars against classical swine fever virus.classical swine fever virus glycoproteins: e2, e(rns) (e0) and e1 are detected on the external part of viral particles and play a major role in the initial stages of viral infection. they form heterodimeric and homodimeric complexes needed to effectively infect host cells. some glycosylation inhibitors, such as tunicamycin, which act at the early stages of glycan chain processing, can influence, not only glycosylation, but also the stability of e2 and e(rns) glycoproteins, effectively inhibiting ...201020153775
phylogenomic analysis of 11 complete african swine fever virus genome sequences.viral molecular epidemiology has traditionally analyzed variation in single genes. whole genome phylogenetic analysis of 123 concatenated genes from 11 asfv genomes, including e75, a newly sequenced virulent isolate from spain, identified two clusters. one contained south african isolates from ticks and warthog, suggesting derivation from a sylvatic transmission cycle. the second contained isolates from west africa and the iberian peninsula. two isolates, from kenya and malawi, were outliers. of ...201020171711
5'-utr-based phylogenetic analysis of classical swine fever virus isolates from india.classical swine fever (csf) caused by classical swine fever virus (csfv) is a globally significant disease of pigs. genetic typing of csfv isolates can help in understanding the epidemiology of disease and trace down the source of outbreak. 5'-utr sequence analysis and subsequent genetic classification of nine csfv field isolates from india indicated that 3 isolates belonged to genotype 2.1 and were closely related to european csfv strains. the remaining 6 isolates belonged to genotype 1 that co ...201020201618
the development of oral fluid-based diagnostics and applications in veterinary medicine.the purpose of this review was to discuss the history of the development and implementation of oral fluid diagnostics for infectious diseases of humans and domestic animals. the use of oral fluid for the assessment of health and diagnosis of disease in humans and animals has a surprisingly long history. as early as 1909, pollaci and ceraulo reported sensitive and specific agglutination of 'micrococcus melitensis' (brucella melitensis) by oral fluid from patients diagnosed with malta fever. a 198 ...201020202287
genetic characterization of e2 gene of classical swine fever virus by restriction fragment length polymorphism and phylogenetic analysis.an rt-nested pcr (rt-npcr)-based restriction fragment length polymorphism (rflp) analyses of the e2 gene were developed for genetic subtyping and differentiation of vaccinated and infected classical swine fever virus (csfv) strains. rt-npcr identified 96 csfv-positive samples from 321 clinical specimens from southeastern china during 2003-2008. the pcr products of positive samples were further differentiated using mspi digestion, 23 were identified as the c-strain, 62 as field strains, and 11 as ...201020217206
patterns of gene expression in swine macrophages infected with classical swine fever virus detected by microarray.infection of domestic swine with highly virulent, classical swine fever virus (csfv) strain brescia, causes lethal disease in all infected animals. however, the molecular mechanisms involved in modulating the host cellular processes and evasion of the immune response have not been clearly established. to gain insight into, the early host response to csfv, we analyzed the pattern of gene expression in infected swine macrophages, using custom designed swine microarrays. macrophages, the target cel ...201020302897
validation of a loop-mediated isothermal amplification assay for visualised detection of wild-type classical swine fever virus.loop-mediated isothermal amplification (lamp) is a nucleic acid amplification method applied and adapted to the detection of a number of pathogens. a lamp assay was developed for the specific detection of wild-type classical swine fever virus (csfv). based on an alignment of genomic sequences of pestiviruses available in genbank, four primers were selected targeting six positions in the ns5b gene region of the viral genome. the assay was performed in a simple water bath at a constant temperature ...201020304010
inhibition of replication of classical swine fever virus in a stable cell line by the viral capsid and staphylococcus aureus nuclease fusion protein.classical swine fever (csf) is one of the major diseases causing serious economic losses to the swine industry. to explore the feasibility of using capsid-targeted viral inactivation (ctvi) as an antiviral strategy against csf infection, a plasmid pcdna-cap-snase was constructed for expressing a fusion protein of csfv capsid (cap) and staphylococcus aureus nuclease (snase). under g418 selection, a mammalian cell line pk-15 expressing stably the fusion protein cap-snase(pk-15/cap-snase) could be ...201020304012
escape of classical swine fever c-strain vaccine virus from detection by c-strain specific real-time rt-pcr caused by a point mutation in the primer-binding site.classical swine fever (csf) is one of the most important diseases of pigs. vaccination in the european union is limited to emergency situations. currently, vaccination for the purpose of disease control is carried out in wild boar populations. wild boar are in most cases vaccinated using an oral bait vaccine based on the live modified vaccine virus c-strain "riems". a real-time reverse transcription polymerase chain reaction (rt-pcr) protocol for differentiation of c-strain "riems" vaccine virus ...201020332004
[preparation and bioactivity of anti-human red blood cell scfv and csfv e2 bifunctional fusion protein].the aim of this study is to construct a bifunctional fusion protein, which can conjugate both human red blood cells and antibodies against classical swine fever virus (csfv). we respectively amplified 2e8scfv and me2 genes from different recombinant vectors, in which 2e8scfv gene is the single chain fv gene against h antigen of human red blood cells, whereas me2 gene is the main antigen coding region gene of csfv e2 protein. we used overlap extension pcr to obtain an artificial fusion gene segme ...201020353089
isolation and characterization of the first chinese strain of porcine teschovirus-8.investigations were carried out to identify the causal agent of acute diarrhea, respiratory distress, and death of pigs on a swine farm in jilin province, northern china. only porcine teschovirus (ptv, designated as ptv-8 jilin/2003) was isolated from samples of organs. the presence of ptv was confirmed by the production of a specific cytopathic effect on susceptible cells and by the results of the immunoperoxidase monolayer assay (ipma), polymerase chain reaction, and electron microscopy. other ...201020362007
an rna-hydrolyzing recombinant antibody exhibits an antiviral activity against classical swine fever virus.some proteins with ribonuclease (rnase) activity have been shown to suppress viral replication. a well-characterized recombinant antibody, 3d8 single-chain variable fragment (3d8 scfv), has rna-hydrolyzing and cell-penetrating activities. here, we investigated antiviral activity of 3d8 scfv against classical swine fever virus (csfv). in a cell line expressing 3d8 scfv (c26), intracellular rna-hydrolysis activity was higher compared to control pk-15 cells and viral replication was strongly suppre ...201020382124
simultaneous detection of porcine circovirus type 2, classical swine fever virus, porcine parvovirus and porcine reproductive and respiratory syndrome virus in pigs by multiplex polymerase chain reaction.a multiplex polymerase chain reaction (pcr) was designed for the simultaneous detection of four viruses involved in reproductive and respiratory failure in pigs: porcine circovirus type 2 (pcv-2), porcine parvovirus (ppv), classical swine fever virus (csfv) and porcine reproductive and respiratory syndrome virus (prrsv). each of the four pairs of oligonucleotide primers exclusively amplified the targeted fragment of the specific viruses. the sensitivity of the multiplex pcr using purified plasmi ...201019131259
cellular processes essential for african swine fever virus to infect and replicate in primary macrophages.the macrophage (mø) is an essential immune cell for innate immunity. such cells are targeted by african swine fever virus (asfv). the early phases of infection with asfv have been previously characterized in non-leukocyte cells such as vero cells. here, we report on several additional key parameters that asfv utilizes during the infection of primary mø. related to virus infection, we established that receptor-mediated endocytosis of the virus by mø is not the exclusive means of entry to infect t ...201019632793
early onset and long lasting protection in pigs provided by a classical swine fever e2-vaccine candidate produced in the milk of goats.for vaccination against classical swine fever virus (csfv), it is strongly desirable to induce a rapid and long lasting protection. at present, only live attenuated csfv vaccines have shown early onset of protection, differing with the recombinant subunit-based vaccines reported so far. recently, a new vaccine formulation based on e2 envelope viral glycoprotein produced in the milk of goats (e2his) has been shown to induce a highly protective response in pigs against csfv infection. pigs immuniz ...201019640591
assessing the impact of climate change on vector-borne viruses in the eu through the elicitation of expert opinion.expert opinion was elicited to undertake a qualitative risk assessment to estimate the current and future risks to the european union (eu) from five vector-borne viruses listed by the world organization for animal health. it was predicted that climate change will increase the risk of incursions of african horse sickness virus (ahsv), crimean-congo haemorrhagic fever virus (cchfv) and rift valley fever virus (rvfv) into the eu from other parts of the world, with african swine fever virus (asfv) a ...201019580695
generation and efficacy evaluation of a recombinant adenovirus expressing the e2 protein of classical swine fever virus.classical swine fever virus (csfv) is the causative agent of classical swine fever (csf), which causes significant economic losses to the pig industry worldwide. the e2 glycoprotein of csfv is the main target for neutralizing antibodies. this study was aimed to develop a recombinant human adenovirus type 5 expressing the csfv e2 gene (radv-e2) and evaluate its efficacy in rabbits and pigs. the results showed that the rabbits and the pigs immunized with the radv-e2 developed high-level csfv-speci ...201019586646
induction of inducible cd4+cd25+foxp3+ regulatory t lymphocytes by porcine reproductive and respiratory syndrome virus (prrsv).increases in numbers or activities of regulatory t lymphocytes (tregs) have been linked to the establishments of several persistent infections. it has been previously shown that porcine reproductive and respiratory syndrome virus (prrsv) can negatively modulate the host immune responses, resulting in persistent infection and secondary immunodeficiency. recently, the existence of porcine cd4(+)cd25(+) tregs has been demonstrated. we investigated the effect of prrsv on the cd4(+)cd25(+) tregs. the ...201019709757
characterization of ns3, ns5a and ns5b of classical swine fever virus through mutation and complementation analysis.in order to get further insight into the organization of the pestiviral replication machinery, characterization of ns3, ns5a and ns5b of classical swine fever virus (csfv) through mutation and complementation analysis was performed. mutation analysis in genomic replicons and subgenomic replicons indicated importance of the gdd motif in ns5b, the deyh motif in ns3 and the conserved sequence c2717-c2740-c2742-c2767 in the ns5a for csfv recover and viral rna synthesis. complementation experiments w ...201019713054
up-regulation of integrin beta3 expression in porcine vascular endothelial cells cultured in vitro by classical swine fever virus.classical swine fever (csf) caused by virulent strains of classical swine fever virus (csfv) is a haemorrhagic disease of pigs, characterized by disseminated intravascular coagulation, thrombocytopenia and immunosuppression. the cell adhesion molecule, integrin beta3, plays a central role in maintaining and regulating vascular permeability. in view of the haemorrhagic pathology of the disease, the effect of csfv infection on integrin beta3 expression was investigated using the swine umbilical ve ...201019767110
rational design of a classical swine fever c-strain vaccine virus that enables the differentiation between infected and vaccinated animals.the c-strain of the classical swine fever virus (csfv) is considered the gold standard vaccine for the control of csf. this vaccine, however, does not enable the serological differentiation between infected and vaccinated animals (diva). consequently, its use can impose severe trade restrictions. the immunodominant and evolutionarily conserved a-domain of the e2 structural glycoprotein is an important target in csfv-specific elisas. with the ultimate aim to render the c-strain suitable as a diva ...201019770004
african swine fever virus polyprotein pp62 is essential for viral core development.one of the most characteristic features of african swine fever virus gene expression is its use of two polyproteins, pp220 and pp62, to produce several structural proteins that account for approximately 32% of the total protein virion mass. equimolecular amounts of these proteins are the major components of the core shell, a thick protein layer that lies beneath the inner envelope, surrounding the viral nucleoid. polyprotein pp220, which is located immediately underneath the internal envelope, i ...201019846532
rna interference targeting nucleocapsid protein (c) inhibits classical swine fever virus replication in sk-6 cells.the application of rna interference (rnai) strategy for controlling classical swine fever could become a promising alternative to the conventional eradication measures, as it was recently shown for foot-and-mouth disease (chen et al., 2004), influenza (ge et al., 2003), porcine reproductive and respiratory syndrome (he et al., 2007) and porcine transmissible gastroenteritis (zhou et al., 2007). the use of synthetic sirna which is corresponding to nucleotides 1130-1148 of the csf virus strain alf ...201019850420
a socio-psychological investigation into limitations and incentives concerning reporting a clinically suspect situation aimed at improving early detection of classical swine fever outbreaks.the aim of this study was to identify limitations and incentives in reporting clinically suspect situations, possibly caused by classical swine fever (csf), to veterinary authorities with the ultimate aim to facilitate early detection of csf outbreaks. focus group sessions were held with policy makers from the veterinary authorities, and representatives of veterinary practitioners and pig farmer unions. personal interviews with a small group of pig farmers and practitioners were held to check li ...201019854004
the effect of tissue degradation on detection of infectious virus and viral rna to diagnose classical swine fever virus.a considerable part of tissue samples that are collected for the monitoring of classical swine fever (csf) from the wild boar population or from domestic pigs are unsuitable for virus detection using the fluorescent antibody test (fat) or virus isolation (vi), due to tissue degradation. reverse transcription polymerase chain reaction (rt-pcr) has a higher sensitivity than the fat and vi, and is supposed to be less sensitive to sample degradation. reliable and quantitative information on how long ...201019854005
characterisation of virus-specific peripheral blood cell cytokine responses following vaccination or infection with classical swine fever viruses.existing live attenuated classical swine fever virus (csfv) vaccines provide a rapid onset of complete protection but pose problems in discriminating infected amongst vaccinated animals. with a view to providing additional information on the cellular mechanisms that may contribute to protection, which in turn may aid the development of the next generation of csfv vaccines, we explored the kinetics of the cytokine responses from peripheral blood cells of pigs vaccinated with an attenuated c-strai ...201019854006
preventive vaccination contributes to control classical swine fever in wild boar (sus scrofa sp.).over the last 20 years, oral vaccination implementing a live attenuated vaccine has been experimented in europe in order to control classical swine fever (csf) in wild boar (sus scrofa sp.). this has generally led to an enhanced seroprevalence and a decreased viroprevalence at the scale of the whole vaccinated populations, but no quantitative analysis has demonstrated the protective effect of preventive vaccination or intensive baiting. in the present paper we conducted a retrospective analysis ...201019854007
a novel combination of taqman rt-pcr and a suspension microarray assay for the detection and species identification of pestiviruses.the genus pestivirus contains four recognized species: classical swine fever virus, border disease virus, bovine viral diarrhoea virus types 1 and 2. all are economically important and globally distributed but classical swine fever is the most serious, concerning losses and control measures. it affects both domestic pigs and wild boars. outbreaks of this disease in domestic pigs call for the most serious measures of disease control, including a stamping out policy in europe. since all the member ...201019854591
in vitro inhibition of the replication of classical swine fever virus by capsid-targeted virus inactivation.classical swine fever virus (csfv) is the causative agent of classical swine fever (csf), a highly contagious fatal disease of swine. few effective antiviral drugs are currently available against csfv infections. to explore the feasibility of using capsid-targeted viral inactivation (ctvi) as an antiviral strategy against csfv infections, we expressed the csfv capsid protein (cap) fused with the nuclease of staphylococcus aureus (sn) in escherichia coli and investigated its effects on the replic ...201019857524
classical swine fever: comparison of oronasal immunisation with cp7e2alf marker and c-strain vaccines in domestic pigs.effective oronasal vaccination against classical swine fever (csf) is essential to achieve protection in wild boar. however the currently available live csf vaccines, e.g. c-strain, do not allow serological differentiation between infected and vaccinated animals (diva). a modified live marker vaccine candidate (cp7e2alf) has been recently developed (reimann et al., 2004). this communication reports the comparison of cp7e2alf and c-strain virus vaccines during 98 days following oronasal immunisat ...201019857935
acute induction of cell death-related ifn stimulated genes (isg) differentiates highly from moderately virulent csfv strains.classical swine fever (csf) severity is dependent on the virulence of the csf virus (csfv) strain. the earliest event detected following csfv infection is a decrease in lymphocytes number. with some csfv strains this leads to lymphopenia, the severity varying according to strain virulence. this lymphocyte depletion is attributed to an induction of apoptosis in non-infected bystander cells. we collected peripheral blood mononuclear cells (pbmc) before and during 3 days post-infection with either ...201019793538
genetic typing of recent classical swine fever isolates from india.seventeen classical swine fever virus (csfv) isolates recovered during the period of 3 years (2006-2008) from india were subjected to nucleotide sequencing in the 5' untranslated region (utr). for genetic typing, 150 nucleotides within this region were used. for better epizootiological understanding, 39 nucleotide sequences of the above region, including 13 indian csfv sequences, available either in the genbank or published literature were also included in the study. based on the phylogenetic an ...201019836905
generation of recombinant pestiviruses using a full-genome amplification strategy.complete genome amplification of viral rna provides a new tool for the generation of modified viruses. we have recently reported a full-genome amplification strategy for recovery of pestiviruses (rasmussen et al., 2008). a full-length cdna amplicon corresponding to the border disease virus-gifhorn genome was generated by long rt-pcr and then rna transcripts derived from this amplicon were used to rescue infectious virus. here, we have now used this full-genome amplification strategy for efficien ...201019836906
classical swine fever in 6- and 11-week-old pigs: haematological and immunological parameters are modulated in pigs with mild clinical disease.the severity of classical swine fever virus (csfv) infection is believed to be determined by different factors, including the virulence of the strain as well as factors related to the host. in the present study, we infected 6- and 11-week-old pigs of unique sanitary status with csfv strain eystrup to elucidate the influence of age on virulence. in both age-groups, a mild clinical course correlated well with the gross-pathological findings at necropsy. the minor variations of clinical, pathologic ...201020709412
control of african swine fever virus replication by small interfering rna targeting the a151r and vp72 genes.african swine fever virus (asfv) is the unique member of the asfarviridae family and asfivirus genus. it is an enveloped double-stranded dna arbovirus that replicates in the cell cytoplasm, similar to poxviruses. there is no vaccine and no treatment available to control this virus.201020710054
peptide affinity chromatography media that bind n(pro) fusion proteins under chaotropic conditions.to design a generic purification platform and to combine the advantages of fusion protein technology and matrix-assisted refolding, a peptide affinity medium was developed that binds inclusion body-derived n(pro) fusion proteins under chaotropic conditions. proteins were expressed in escherichia coli using an expression system comprising the autoprotease n(pro) from pestivirus, or its engineered mutant called eddie, with c-terminally linked target proteins. upon refolding, the autoprotease becam ...201020800233
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