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effectiveness of the e2-classical swine fever virus recombinant vaccine produced and formulated within whey from genetically transformed goats.subunit recombinant vaccines against classical swine fever virus (csfv) are a promising alternative to overcome practical and biosafety issues with inactivated vaccines. one of the strategies in evaluation under field conditions is the use of a new marker e2-based vaccine produced in the milk of adenovirally transduced goats. previously we had demonstrated the efficacy of this antigen, which conferred early protection and long-lasting immunity in swine against csfv infection. here, we have used ...201425274802
efficacy of csf vaccine cp7_e2alf in piglets with maternally derived antibodies.there is a need for live diva (differentiating infected from vaccinated animals) vaccines against classical swine fever (csf). the aim of this study was to investigate whether vaccination with the chimeric pestivirus vaccine cp7_e2alf is efficacious to protect young piglets born from vaccinated sows, thus with maternally derived antibodies (mdas). groups of 10 piglets each, with or without mdas, were vaccinated either intramuscularly (im), at an age of 3 or 6 weeks, or orally (or), at an age of ...201425265929
dynamics of african swine fever virus shedding and excretion in domestic pigs infected by intramuscular inoculation and contact transmission.african swine fever virus (asfv) is a highly virulent swine pathogen that has spread across eastern europe since 2007 and for which there is no effective vaccine or treatment available. the dynamics of shedding and excretion is not well known for this currently circulating asfv strain. therefore, susceptible pigs were exposed to pigs intramuscularly infected with the georgia 2007/1 asfv strain to measure those dynamics through within- and between-pen transmission scenarios. blood, oral, nasal an ...201425256695
rescue of the highly virulent classical swine fever virus strain "koslov" from cloned cdna and first insights into genome variations relevant for virulence.classical swine fever virus (csfv) strain "koslov" is highly virulent with a mortality rate of up to 100% in pigs. in this study, we modified non-functional cdnas generated from the blood of koslov virus infected pigs by site-directed mutagenesis, removing non-synonymous mutations step-by-step, thereby producing genomes encoding the consensus amino acid sequence. viruses rescued from the construct corresponding to the inferred parental form were highly virulent, when tested in pigs, with infecte ...201425240324
evaluation of hemostaseological status of pigs experimentally infected with african swine fever virus.african swine fever is a highly contagious hemorrhagic disease of pigs caused by african swine fever virus (asfv). hemorrhages are the most frequently reported lesions in acute and subacute forms of asf. hemorrhagic lesions are accompanied by impaired hemostasis, which includes thrombocytopenia and changes in the coagulation system. in the present study, experimental infection was conducted to elucidate whether a highly virulent asfv genotype ii circulating in the trans-caucasus and eastern euro ...201425239678
characterization of the c-terminal sequence of ns5a necessary for the assembly and production of classical swine fever virus infectious particles.recent studies show that classical swine fever virus (csfv) ns5a is an essential replicase component, but it is not known how ns5a participates in viral particle production. in this study, deletion and substitution mutations were introduced into the c-terminus of csfv ns5a. the efficiency of core protein release and extracellular and intracellular infectivity levels were assessed and ns5a-core interaction was investigated. these results suggested that csfv ns5a was a key factor for the assembly ...201425218811
expression library immunization can confer protection against lethal challenge with african swine fever virus.african swine fever is one of the most devastating pig diseases, against which there is no vaccine available. recent work from our laboratory has demonstrated the protective potential of dna vaccines encoding three african swine fever viral antigens (p54, p30, and the hemagglutinin extracellular domain) fused to ubiquitin. partial protection was afforded in the absence of detectable antibodies prior to virus challenge, and survival correlated with the presence of a large number of hemagglutinin- ...201425210179
generation and efficacy evaluation of recombinant classical swine fever virus e2 glycoprotein expressed in stable transgenic mammalian cell line.classical swine fever virus (csfv) is the causative agent of classical swine fever (csf), which is a highly contagious swine disease that causes significant economic loses to the pig industry worldwide. the envelope e2 glycoprotein of csfv is the most important viral antigen in inducing protective immune response against csf. in this study, we generated a mammalian cell clone (bcsfv-e2) that could stably produce a secreted form of csfv e2 protein (me2). the me2 protein was shown to be n-linked g ...201425198669
extensive homologous recombination in classical swine fever virus: a re-evaluation of homologous recombination events in the strain af407339.in this short report, the genome-wide homologous recombination events were re-evaluated for classical swine fever virus (csfv) strain af407339. we challenged a previous study which suggested only one recombination event in af407339 based on 25 csfv genomes. through our re-analysis on the 25 genomes in the previous study and the 41 genomes used in the present study, we argued that there should be possibly at least two clear recombination events happening in af407339 through genome-wide scanning. ...201425183941
up-regulated expression of pd-1 and its ligands during acute classical swine fever virus infection in swine.in order to investigate the relationship between the pd-1 pathway and impairment of immune responses with the csfv infection, the mrna expression of pd-1 and its ligands were evaluated by quantitative polymerase chain reaction (qpcr) during artificial csfv infection. simultaneously, expression of il-2 and il-10 mrna were detected. the t cell proliferation and csfv load in plasma were also measured. results showed that the expression of pd-1 and its ligands mrna were significantly increased (p < ...201425178664
genetic variation among african swine fever genotype ii viruses, eastern and central europe.african swine fever virus (asfv) was first reported in eastern europe/eurasia in 2007. continued spread of asfv has placed central european countries at risk, and in 2014, asfv was detected in lithuania and poland. sequencing showed the isolates are identical to a 2013 asfv from belarus but differ from asfv isolated in georgia in 2007.201425148518
classical swine fever virus and p7 protein induce secretion of il-1β in macrophages.classical swine fever virus (csfv) has a tropism for vascular endothelial cells and immune system cells. the process and release of pro-inflammatory cytokines, including il-1β and il-18, is one of the fundamental reactions of the innate immune response to viral infection. in this study, we investigated the production of il-1β from macrophages following csfv infection. our results showed that il-1β was upregulated after csfv infection through activating caspase-1. subsequent studies demonstrated ...201425146005
predominance of genotype 1.1 and emergence of genotype 2.2 classical swine fever viruses in north-eastern region of india.classical swine fever (csf) is a highly contagious and the most important disease of pigs worldwide.csf is enzootic in pig herds in india and continues to cause huge economic losses to pig farmers. nearly 40% of the total pig population of india is present in the north-eastern (ne) states where pig husbandry plays an important role in the socio-economic development. pigs reared in the backyards are the only source of livelihood for a majority of poor tribal population in the region. hardly any c ...201425135466
establishment and application of a multiplex pcr for rapid and simultaneous detection of six viruses in swine.a multiplex pcr assay was developed and evaluated subsequently for its effectiveness in simultaneously detecting mixed viral infections of swine. specific primers were designed and used for testing the six swine viruses: three dna viruses, including pseudorabies virus (prv), porcine parvovirus (ppv), and porcine circovirus type 2 (pcv2); three common rna viruses, including porcine reproductive and respiratory syndrome virus (prrsv), classical swine fever virus (csfv), and japanese encephalitis v ...201425116201
development of an evagreen-based multiplex real-time pcr assay with melting curve analysis for simultaneous detection and differentiation of six viral pathogens of porcine reproductive and respiratory disorder.concurrent infection of pigs with two or more pathogens is common in pigs under intensive rearing conditions. porcine circovirus type 2 (pcv2), porcine parvovirus (ppv), porcine reproductive and respiratory syndrome virus (prrsv), classical swine fever virus (csfv), japanese encephalitis virus (jev) and pseudorabies virus (prv) are all associated with reproductive or respiratory disorders or both and can cause significant economic losses in pig production worldwide. an evagreen-based multiplex r ...201425102430
induction of immune responses in mice and pigs by oral administration of classical swine fever virus e2 protein expressed in rice calli.classical swine fever (csf), caused by the csf virus (csfv), is a highly contagious disease in pigs. in korea, vaccination using a live-attenuated strain (lom strain) has been used to control the disease. however, parenteral vaccination using a live-attenuated strain still faces a number of problems related to storage, cost, injection stress, and differentiation of csfv infected and vaccinated pigs. therefore, two kinds of new candidates for oral vaccination have been developed based on the tran ...201425091740
seroprevalence and associated risk factors of important pig viral diseases in bhutan.a cross-sectional serological study was conducted in bhutan between october 2011 and february 2012 to determine the prevalence of antibodies to classical swine fever virus (csfv), porcine reproductive and respiratory syndrome virus (prrsv), porcine circovirus type 2 (pcv2), swine influenza virus (siv) subtype h1n1 and aujeszky's disease virus (adv). furthermore, risk factors for the seropositive status were investigated. antibodies to siv, subtype h1n1 (likely pandemic h1n1 2009) were detected i ...201425081946
partial activation of natural killer and γδ t cells by classical swine fever viruses is associated with type i interferon elicited from plasmacytoid dendritic cells.vaccination with live attenuated classical swine fever virus (csfv) vaccines can rapidly confer protection in the absence of neutralizing antibodies. with an aim of providing information on the cellular mechanisms that may mediate this protection, we explored the interaction of porcine natural killer (nk) cells and γδ t cells with csfv. both nk and γδ t cells were refractory to infection with attenuated or virulent csfv, and no stimulatory effects, as assessed by the expression of major histocom ...201425080554
vaccine potential of two previously uncharacterized african swine fever virus isolates from southern africa and heterologous cross protection of an avirulent european isolate.african swine fever (asf) is a mostly fatal viral infection of domestic pigs for which there is no vaccine available. the disease is endemic to most of sub-saharan africa, causes severe losses and threatens food security in large parts of the continent. naturally occurring attenuated asf viruses have been tested as vaccine candidates, but protection was variable depending on the challenge virus. in this study, the virulence of two african isolates, one from a tick vector and the other from an in ...201625073549
comparative analyses of host responses upon infection with moderately virulent classical swine fever virus in domestic pigs and wild boar.classical swine fever (csf) is one of the most important viral diseases of pigs. clinical signs may vary from almost inapparent infection to a hemorrhagic fever like illness. among the host factors leading to different disease courses are age, breed, and immune status. the aim of this study was to compare host responses of different pig breeds upon infection with a recent moderately virulent csf virus (csfv) strain, and to assess their impact on the clinical outcome and the efficiency of immune ...201425073480
drivers and risk factors for circulating african swine fever virus in uganda, 2012-2013.we explored observed risk factors and drivers of infection possibly associated with african swine fever (asf) epidemiology in uganda. representative sub-populations of pig farms and statistics were used in a case-control model. indiscriminate disposal of pig viscera and waste materials after slaughter, including on open refuse dumps, farm-gate buyers collecting pigs and pig products from within a farm, and retention of survivor pigs were plausible risk factors. wire mesh-protected windows in pig ...201425066802
tissues expression, polymorphisms identification of fcrn gene and its relationship with serum classical swine fever virus antibody level in pigs.neonatal fc receptor (fcrn) gene encodes a receptor that binds the fc region of monomeric immunoglobulin g (igg) and is responsible for igg transport and stabilization. in this report, the 8,900 bp porcine fcrn genomic dna structure was identified and putative fcrn protein included 356 amino acids. alignment and phylogenetic analysis of the porcine fcrn amino acid sequences with their homologies of other species showed high identity. tissues expression of fcrn mrna was detected by real time quan ...025049667
a sensitive multiplex real-time pcr panel for rapid diagnosis of viruses associated with porcine respiratory and reproductive disorders.the objective of this study was to develop a multiplex real-time pcr panel using taqman probes for the detection and differentiation of porcine circovirus type 2 (pcv2), porcine reproductive and respiratory syndrome virus north american type (prrsv-na), pseudorabies virus (prv), classical swine fever virus (csfv), porcine parvovirus type 1 (ppv1) and japanese encephalitis virus (jev). specific primer and probe combinations for pcv2, prrsv, prv, csfv, ppv1 and jev were selected within the conserv ...201425044282
comparison of conventional rt-pcr, reverse-transcription loop-mediated isothermal amplification, and sybr green i-based real-time rt-pcr in the rapid detection of bovine viral diarrhea virus nucleotide in contaminated commercial bovine sera batches.bovine viral diarrhea virus (bvdv) can contaminate biological products produced in bovine or porcine cells or manufactured using bovine sera. a rapid, specific, sensitive, and practical method of detecting bvdv in bio-products is needed. the purpose of this study was to compare three assays with respect to their ability to accurately detect bvdv in biological samples, namely reverse-transcription loop-mediated isothermal amplification (rt-lamp), sybr green i-based real-time rt-pcr, and conventio ...201425019170
suitability of faeces and tissue samples as a basis for non-invasive sampling for african swine fever in wild boar.a challenging aspect of asfv control in wild boar populations is the design and implementation of effective surveillance and monitoring programmes, both for early warning, and to determine the ongoing epidemiological situation in an infected population. testing blood samples requires invasive sampling strategies like hunting or capture of wild boar. besides being biased towards healthy animals, such strategies are also linked to further spread of the virus. non-invasive sampling strategies would ...201425017975
interaction of structural core protein of classical swine fever virus with endoplasmic reticulum-associated degradation pathway protein os9.classical swine fever virus (csfv) core protein is involved in virus rna protection, transcription regulation and virus virulence. to discover additional core protein functions a yeast two-hybrid system was used to identify host proteins that interact with core. among the identified host proteins, the osteosarcoma amplified 9 protein (os9) was further studied. using alanine scanning mutagenesis, the os9 binding site in the csfv core protein was identified, between core residues (90)iaim(93), nea ...201425010283
molecular characterization of african swine fever virus from domestic pigs in northern tanzania during an outbreak in 2013.african swine fever (asf) is an acute, highly contagious and deadly viral hemorrhagic fever of domestic pigs caused by african swine fever virus (asfv), a double-stranded dna virus of the family asfarviridae. in this study, molecular diagnosis and characterization of outbreak asfv in northern tanzania, was performed on spleen, lymph node, kidney, and heart samples collected in june and july 2013 from domestic pigs that died during a hemorrhagic disease outbreak. confirmatory diagnosis of asf was ...201424996815
modular framework to assess the risk of african swine fever virus entry into the european union.the recent occurrence and spread of african swine fever (asf) in eastern europe is perceived as a serious risk for the pig industry in the european union (eu). in order to estimate the potential risk of asf virus (asfv) entering the eu, several pathways of introduction were previously assessed separately. the present work aimed to integrate five of these assessments (legal imports of pigs, legal imports of products, illegal imports of products, fomites associated with transport and wild boar mov ...201424992824
no evidence of african swine fever virus replication in hard ticks.african swine fever (asf) is caused by african swine fever virus (asfv), a tick-borne dna virus. soft ticks of the genus ornithodoros are the only biological vectors of asfv recognized so far. although other hard ticks have been tested for vector competence, two commonly found tick species in europe, ixodes ricinus and dermacentor reticulatus, have not been assessed for their vector competence for asfv. in this study, we aimed to determine whether virus replication can occur in any of these two ...201424980962
specific ligands for classical swine fever virus screened from landscape phage display library.classical swine fever (csf) is a devastating infectious disease caused by classical swine fever virus (csfv). the screening of csfv-specific ligands is of great significance for diagnosis and treatment of csf. affinity selection from random peptide libraries is an efficient approach to discover ligands with high stability and specificity. here, we screened phage ligands for the csfv e2 protein from f8/8 landscape phage display library by biopanning and obtained four phage clones specific for the ...201424977927
generation of a recombinant classical swine fever virus stably expressing the firefly luciferase gene for quantitative antiviral assay.classical swine fever (csf), caused by classical swine fever virus (csfv), is a highly contagious swine disease leading to significant economic losses worldwide. vaccines are widely used to control the disease, and no csfv-specific antivirals are currently available. to facilitate anti-csfv molecule discovery, we developed a reporter virus csfv-n(pro)fluc stably expressing the firefly luciferase (fluc) gene in the n(pro) gene. the reporter virus enabled more sensitive and convenient detection of ...201424956495
effect of specific amino acid substitutions in the putative fusion peptide of structural glycoprotein e2 on classical swine fever virus replication.e2, along with e(rns) and e1, is an envelope glycoprotein of classical swine fever virus (csfv). e2 is involved in several virus functions: cell attachment, host range susceptibility and virulence in natural hosts. here we evaluate the role of a specific e2 region, (818)cpigwtgviec(828), containing a putative fusion peptide (fp) sequence. reverse genetics utilizing a full-length infectious clone of the highly virulent csfv strain brescia (bicv) was used to evaluate how individual amino acid subs ...201424889231
modern adjuvants do not enhance the efficacy of an inactivated african swine fever virus vaccine preparation.african swine fever (asf) is among the most devastating viral diseases of pigs. in recent years, the disease has spread alarmingly. despite intensive research activities, promising vaccine candidates are still lacking. for this reason, a study was undertaken to re-assess inactivated asfv preparations with state-of-the-art adjuvants. inactivated preparations of asf virus (asfv) "armenia08" were adjuvanted with either polygen™ or emulsigen(®)-d, respectively, and used to immunize six weaner pigs t ...201424877766
complete genome sequence of classical swine fever virus genotype 2.2 strain bergen.the complete genome sequence of the genotype 2.2 classical swine fever virus strain bergen has been determined; this strain was originally isolated from persistently infected domestic pigs in the netherlands and is characterized to be of low virulence.201424874676
preclinical detection of porcine circovirus type 2 infection using an ultrasensitive nanoparticle dna probe-based pcr assay.porcine circovirus type 2 (pcv2) has emerged as one of the most important pathogens affecting swine production globally. preclinical identification of pcv2 is very important for effective prophylaxis of pcv2-associated diseases. in this study, we developed an ultrasensitive nanoparticle dna probe-based pcr assay (undp-pcr) for pcv2 detection. magnetic microparticles coated with pcv2 specific dna probes were used to enrich pcv2 dna from samples, then gold nanoparticles coated with pcv2 specific o ...201424842840
the classic swine fever virus (csfv) core protein can enhance de novo-initiated rna synthesis by the csfv polymerase ns5b.study of the classical swine fever virus (csfv) replication is challenging because it is a bsl-3-ag pathogen that requires specialized facilities. we developed a cell-based assay in human embryonic kidney 293t cells that can quantify the activities of ns5b, the csfv rna-dependent rna polymerase. the 5br assay uses transiently-expressed csfv ns5b to produce rnas that activate the rig-i-mediated signaling pathway to result in reporter protein production. upon co-expression of the csfv core protein ...201424824302
visualization of the npro protein in living cells using biarsenically labeling tetracysteine-tagged classical swine fever virus.real-time fluorescence imaging of viral proteins in living cells is a valuable means to study virus-host interactions, and tetracysteine (tc)-biarsenical technology has been used in several viruses but not in classical swine fever virus (csfv). here, we generated csfv mutants vsmtc385 or vsmtc412 bearing the small tc tag (ccpgcc) in the n-terminal region of the n(pro) protein. the mutants showed growth characteristics indistinguishable from that of the wild-type virus, and retained similar n(pro ...201424815879
whole-genome sequence of a classical swine fever virus isolated from the uttarakhand state of india.we report the first complete genome sequence of a classical swine fever (csf) virus of subgenotype 2.2. the virus (csfv/ind/uk/lal-290) was isolated from the uttarakhand state of india from a backyard pig suspected of having csf. this genome sequence will give useful insight for future molecular epidemiological studies and the development of an effective vaccine in india.201424812219
early lethality of shrna-transgenic pigs due to saturation of microrna pathways.rna interference (rnai) is considered as a potential modality for clinical treatment and anti-virus animal breeding. here, we investigate the feasibility of inhibiting classical swine fever virus (csfv) replication by short hairpin rna (shrna) in vitro and in vivo. we generate four different shrna-positive clonal cells and two types of shrna-transgenic pigs. csfv could be effectively inhibited in shrna-positive clonal cells and tail tip fibroblasts of shrna-transgenic pigs. unexpectedly, an earl ...201424793764
a novel bromodeoxyuridine-resistant wild boar lung cell line facilitates generation of african swine fever virus recombinants.manipulation of african swine fever virus (asfv) genomes, in particular those from field strains, is still a challenge. we have shown recently that generation of a green-fluorescent-protein-expressing, thymidine-kinase-negative (tk-) mutant of the low-pathogenic african swine fever virus field strain nhv was supported by a tk- vero cell line. since nhv, like other asfv field strains, does not replicate well in vero cells, a bromodeoxyuridine (brdu)- resistant cell line derived from wild boar lun ...201424793370
classical swine fever in china: a minireview.classical swine fever (csf), caused by classical swine fever virus (csfv), is an oie-listed, highly contagious, often fatal disease of swine worldwide. currently, the disease is controlled by prophylactic vaccination in china and many other countries using the modified live vaccines derived from c-strain, which was developed in china in the mid-1950s. this minireview summarizes the epidemiology, diagnostic assays, control and challenges of csf in china. though csf is essentially under control, c ...201424793098
discovering up-regulated vegf-c expression in swine umbilical vein endothelial cells by classical swine fever virus shimen.infection of domestic swine with the highly virulent shimen strain of classical swine fever virus causes hemorrhagic lymphadenitis and diffuse hemorrhaging in infected swine. we analyzed patterns of gene expression for csfv shimen in swine umbilical vein endothelial cells (suvecs). transcription of the vascular endothelial growth factor (vegf) c gene (vegf-c) and translation of the corresponding protein were significantly up-regulated in suvecs. our findings suggest that vegf-c is involved in me ...201424758593
arboviruses pathogenic for domestic and wild animals.the objective of this chapter is to provide an updated and concise systematic review on taxonomy, history, arthropod vectors, vertebrate hosts, animal disease, and geographic distribution of all arboviruses known to date to cause disease in homeotherm (endotherm) vertebrates, except those affecting exclusively man. fifty arboviruses pathogenic for animals have been documented worldwide, belonging to seven families: togaviridae (mosquito-borne eastern, western, and venezuelan equine encephaliliti ...201424751197
vaccination influences the evolution of classical swine fever virus.classical swine fever is a serious, economically damaging disease caused by classical swine fever virus (csfv). the csfv is composed of two clades, according to phylogenetic estimates. attenuated live vaccine such as hclv, has been widely used to protect pigs from csfv, but the influence of vaccination on the evolution of csfv has not been studied. we conducted a systemic analysis of the impact of vaccination on the evolution of csfv by comparing vaccine-related and non-vaccine-related csfv grou ...201424747964
npro of classical swine fever virus contributes to pathogenicity in pigs by preventing type i interferon induction at local replication sites.classical swine fever (csf) caused by csf virus (csfv) is a highly contagious disease of pigs. the viral protein npro of csfv interferes with alpha- and beta-interferon (ifn-α/β) induction by promoting the degradation of interferon regulatory factor 3 (irf3). during the establishment of the live attenuated csf vaccine strain gpe-, npro acquired a mutation that abolished its capacity to bind and degrade irf3, rendering it unable to prevent ifn-α/β induction. in a previous study, we showed that th ...201424742209
first isolation of border disease virus in japan is from a pig farm with no ruminants.the first isolation of border disease virus (bdv) in japan was from a pig farm of the farrow-to-finishing type that kept no small ruminants or cattle. the infection was detected in the course of sero-surveillance for classical swine fever virus (csfv) in japan. the infected pigs had no clinical symptoms of csfv or other disease; nevertheless, a high sero-positive rate of 58.5% was identified. a persistently infected pig with the bdv was found and suspected to be the cause of sero-prevalence in t ...201424735918
clinical presentation resembling mucosal disease associated with 'hobi'-like pestivirus in a field outbreak.the genus pestivirus of the family flaviviridae consists of four recognized species: bovine viral diarrhoea virus 1 (bvdv-1), bovine viral diarrhoea virus 2 (bvdv-2), classical swine fever virus (csfv) and border disease virus (bdv). recently, atypical pestiviruses ('hobi'-like pestiviruses) were identified in batches of contaminated foetal calf serum and in naturally infected cattle with and without clinical symptoms. here, we describe the first report of a mucosal disease-like clinical present ...201624735072
synthesis and antiviral activity of a novel glycosyl sulfoxide against classical swine fever virus.a novel compound-2″,3″,4″,6″-tetra-o-acetyl-β-d-galactopyranosyl-(1→4)-2',3',6'-tri-o-acetyl-1-thio-β-d-glucopyranosyl-(5-nitro-2-pyridyl) sulfoxide-designated gp6 was synthesized and assayed for cytotoxicity and in vitro antiviral properties against classical swine fever virus (csfv) in this study. we showed that the examined compound effectively arrested csfv growth in swine kidney cells (sk6) at a 50% inhibitory concentration (ic50) of 5 ± 0.12 μg/ml without significant toxicity for mammalian ...201424721828
detection and genetic analysis of porcine bocavirus in different swine herds in north central china.porcine bocavirus (pbov) has been reported to be associated with postweaning multisystemic wasting syndrome and pneumonia in pigs. in this study, a survey was conducted to evaluate the prevalence of pbov in slaughter pigs, sick pigs, asymptomatic pigs and classical swine fever virus (csfv) eradication plan herds in five provinces of china (henan, liaoning, shandong, hebei and tianjin) by means of pcr targeting ns1 gene of pbov. among the total of 403 tissue samples, 11.41% were positive for pbov ...201424701194
the csfv dnachip: a novel diagnostic assay for classical swine fever virus.a novel assay, the csfv dnachip, was developed to clearly and rapidly discriminate three genotypes of classical swine fever virus (csfv). total rna was extracted from clinical samples and then subjected to a one-step reverse-transcription polymerase chain reaction (rt-pcr) using cy3-labeled primers from the 5' non-coding region (ncr) of csfv. amplicons were hybridized to the csfv dnachip and fluorescence scanning was performed for detection of csfv. a cut-off fluorescence intensity value of 5000 ...201424698761
substituted 2,6-bis(benzimidazol-2-yl)pyridines: a novel chemical class of pestivirus inhibitors that targets a hot spot for inhibition of pestivirus replication in the rna-dependent rna polymerase.2,6-bis(benzimidazol-2-yl)pyridine (bbp/csfa-0) was identified in a cpe-based screening as a selective inhibitor of the in vitro bovine viral diarrhea virus (bvdv) replication. the ec50-values for the inhibition of bvdv-induced cytopathic (cpe) effect, viral rna synthesis and the production of infectious virus were 0.3±0.1μm, 0.05±0.01μm and 0.3±0.04μm, respectively. furthermore, bbp/csfa-0 inhibits the in vitro replication of the classical swine fever virus (csfv) with an ec50 of 0.33±0.25μm. b ...201424680957
identification and epidemiology of a rare hobi-like pestivirus strain in bangladesh.the genus pestivirus of the family flaviviridae consists of four recognized species: bovine viral diarrhoea virus 1 (bvdv-1), bovine viral diarrhoea virus 2 (bvdv-2), classical swine fever virus and border disease virus. a new putative pestivirus species tentatively named as either 'hobi-like pestivirus' or bvdv-3 has recently been identified in brazil, italy and thailand. despite reports of serological evidence of bvdv in bangladesh, the types of the virus circulating in cattle have not been id ...201424650238
a new nanopcr molecular assay for detection of porcine bocavirus.nanoparticle-assisted polymerase chain reaction (nanopcr) is a novel method for the rapid amplification of dna and has been used for the detection of virus. for detection of porcine bocavirus (pbov), a sensitive and specific nanopcr assay was developed with a pair of primers that were designed based on ns1 gene sequences available in genbank. under the optimized conditions of the pbov nanopcr assay, the nanopcr assay was 100-fold more sensitive than a conventional pcr assay. the lower detection ...201424642242
diva vaccine properties of the live chimeric pestivirus strain cp7_e2gif.live modified vaccines to protect against classical swine fever virus (csfv), based on chimeric pestiviruses, have been developed to enable serological differentiation of infected from vaccinated animals (diva). in this context, the chimeric virus cp7_e2gif vaccine candidate is unique as it does not include any csfv components. in the present study, the diva vaccine properties of cp7_e2gif were evaluated in comparison to the conventional live attenuated riemser c-strain vaccine. sera and tonsil ...201424629779
how a low-fidelity dna polymerase chooses non-watson-crick from watson-crick incorporation.a dogma for dna polymerase catalysis is that the enzyme binds dna first, followed by mgdntp. this mechanism contributes to the selection of correct dntp by watson-crick base pairing, but it cannot explain how low-fidelity dna polymerases overcome watson-crick base pairing to catalyze non-watson-crick dntp incorporation. dna polymerase x from the deadly african swine fever virus (pol x) is a half-sized repair polymerase that catalyzes efficient dg:dgtp incorporation in addition to correct repair. ...201424617852
autocatalytic activity and substrate specificity of the pestivirus n-terminal protease npro.pestivirus n(pro) is the first protein translated in the viral polypeptide, and cleaves itself off co-translationally generating the n-terminus of the core protein. once released, n(pro) blocks the host׳s interferon response by inducing degradation of interferon regulatory factor-3. n(pro׳)s intracellular autocatalytic activity and lack of trans-activity have hampered in vitro cleavage studies to establish its substrate specificity and the roles of individual residues. we constructed n(pro)-gfp ...201424606708
first assessment of classical swine fever marker vaccine candidate cp7_e2alf for oral immunization of wild boar under field conditions.oral vaccination against classical swine fever (csf) is a potent tool to control disease outbreaks in wild boar. so far, vaccination campaigns have been carried out using live attenuated vaccines that do not allow serological differentiation of infected from vaccinated animals (diva). although this drawback is acceptable for wild boar, the use of marker vaccines would facilitate studies on disease and vaccination dynamics. recently, the csf marker vaccine candidate cp7_e2alf was assessed for ora ...201424565752
the role of noncoding regions of classical swine fever virus c-strain in its adaptation to the rabbit.classical swine fever (csf) is a devastating disease of swine caused by classical swine fever virus (csfv). c-strain, a modified live vaccine against csf, was developed through hundreds of passages of a highly virulent csfv in the rabbit in china in the mid-1950s. to identify the role of noncoding regions (ncrs) of c-strain in its adaptation to the rabbit, we generated and evaluated a series of chimeric viruses derived from c-strain and the highly virulent shimen strain. the results demonstrated ...201424553098
mapping quantitative trait loci for the lysozyme level and immunoglobulin g blocking percentage of classical swine fever virus.increased disease resistance through improved general immune capacity would be beneficial for the welfare and productivity of farm animals. classical swine fever (csf) is a contagious disease in farm animals. the immunoglobulin g (igg) blocking percentage of csf virus (csfv) in serum is an essential diagnostic parameter in veterinary practice. in addition, lysozymes are a part of the innate immune system. to identify quantitative trait loci (qtl) for igg blocking percentage of csfv and lysozyme ...201424535855
the untranslated regions of classic swine fever virus rna trigger apoptosis.classical swine fever virus (csfv) causes a broad range of disease in pigs, from acute symptoms including high fever and hemorrhages, to chronic disease or unapparent infection, depending on the virus strain. csfv belongs to the genus pestivirus of the family flaviviridae. it carries a single-stranded positive-sense rna genome. an internal ribosomal entry site (ires) in the 5' untranslated region (utr) drives the translation of a single open reading frame encoding a 3898 amino acid long polypept ...201424533157
cytokine and immunoglobulin isotype profiles during cp7_e2alf vaccination against a challenge with the highly virulent koslov strain of classical swine fever virus.cp7_e2alf is a promising marker vaccine candidate against classical swine fever (csf). to better understand the mechanisms of protection, cytokine and isotype-specific antibody profiles were investigated in cp7_e2alf vaccinated pigs before and after challenge with the highly virulent csfv strain "koslov" at 14 days or 6 months post-vaccination. the interference of vaccination with csfv pathogeny-related cytokine responses, previously described following a moderately virulent challenge, was confi ...201424530018
in vitro inhibition of the replication of classical swine fever virus by porcine mx1 protein.classical swine fever virus (csfv) is the causative pathogen of classical swine fever (csf), a highly contagious disease of swine. mx proteins are interferon-induced dynamin-like gtpases present in all vertebrates with a wide range of antiviral activities. although zhao et al. (2011) have reported that human mxa can inhibit csfv replication, whether porcine mx1 (pomx1) has anti-csfv activity remains unknown. in this study, we generated a cell line designated pk-15/egfp-pomx1 which expressed porc ...201424500530
screening of cellular proteins that interact with the classical swine fever virus non-structural protein 5a by yeast two-hybrid analysis.classical swine fever virus (csfv), the pathogen of classical swine fever (csf), causes severe hemorrhagic fever and vascular necrosis in domestic pigs and wild boar. a large number of evidence has proven that non-structural 5a (ns5a) is not only a very important part of viral replication complex, but also can regulate host cell's function; however, the underlying mechanisms remain poorly understood. in the current study, aiming to find more clues in understanding the molecular mechanisms of csf ...201424499791
enhancing expression of the classical swine fever virus glycoprotein e2 in yeast and its application to a blocking elisa.classical swine fever virus (csfv) infection is a severe swine disease, often causing large economic losses. a pichia pastoris yeast-expressed csfv glycoprotein e2 (ye2) has been shown to induce a protective immune response against the virus. to improve the expression level of ye2, the first codon of e2 gene, arg (cgg), which is the least used in p. pastoris, was optimized to the most favorite codon aga. the yield of e2 protein was remarkably increased in the codon optimized strain (n342). three ...201424468422
development of colloidal gold-based immunochromatographic assay for rapid detection of mycoplasma suis in porcine plasma.a one-step immunochromatographic assay using gold nanoparticles coated with polyclonal antibody (pab) against mycoplasma suis (m. suis) was developed in this study for the detection of m. suis in porcine plasma. the colloidal gold was prepared by the reduction of gold salt with sodium citrate coupled with pab against m. suis. the pab was produced by immunizing the balb/c mice with recombinant msg1 (rmsg1) protein from m. suis expressed in escherichia coli. the optimal concentrations of the captu ...201424434494
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
interaction of csfv e2 protein with swine host factors as detected by yeast two-hybrid system.e2 is one of the envelope glycoproteins of pestiviruses, including classical swine fever virus (csfv) and bovine viral diarrhea virus (bvdv). e2 is involved in several critical functions, including virus entry into target cells, induction of a protective immune response and virulence in swine. however, there is no information regarding any host binding partners for the e2 proteins. here, we utilized the yeast two-hybrid system and identified fifty-seven host proteins as positive binding partners ...201424416391
efficacy of marker vaccine candidate cp7_e2alf against challenge with classical swine fever virus isolates of different genotypes.classical swine fever (csf) is among the most important viral disease of domestic and feral pigs and has a serious impact on animal health and pig industry. in most countries with industrialized pig production, prophylactic vaccination against csf is banned, and all efforts are directed towards eradication of the disease, e.g. by culling of infected herds and animal movement restrictions. nevertheless, emergency vaccination remains an option to minimize the socio-economic impact of outbreaks. fo ...201424411658
correlation of cell surface marker expression with african swine fever virus infection.the expression of surface markers on african swine fever virus (asfv) infected cells was evaluated to assess their involvement in infection. previous findings indicated cd163 expression was correlated with asfv susceptibility. however, in this study the expression of porcine cd163 on cell lines did not increase the infection rate of these cells indicating other factors are likely to be important in determining susceptibility to infection. on adherent porcine bone marrow (pbm) cells the expressio ...201424398227
n pro of bungowannah virus exhibits the same antagonistic function in the ifn induction pathway than that of other classical pestiviruses.bungowannah virus is the most divergent atypical pestivirus that had been detected up to now, and does not fit into any of the four approved species: bovine viral diarrhea virus type 1 (bvdv-1) and type 2 (bvdv-2), classical swine fever virus (csfv) and border disease virus (bdv). however, the presence of n(pro) and e(rns) coding regions, which are unique to pestiviruses, provides clear evidence of a pestivirus. nevertheless, the amino acid identity of bungowannah virus n(pro) and bvdv-1 n(pro) ...201424398226
proteome-wide screening reveals immunodominance in the cd8 t cell response against classical swine fever virus with antigen-specificity dependent on mhc class i haplotype expression.vaccination with live attenuated classical swine fever virus (csfv) vaccines induces a rapid onset of protection which has been associated with virus-specific cd8 t cell ifn-γ responses. in this study, we assessed the specificity of this response, by screening a peptide library spanning the csfv c-strain vaccine polyprotein to identify and characterise cd8 t cell epitopes. synthetic peptides were pooled to represent each of the 12 csfv proteins and used to stimulate pbmc from four pigs rendered ...201324376799
porcine circovirus type 2 decreases the infection and replication of attenuated classical swine fever virus in porcine alveolar macrophages.recently, it has been noted that porcine circovirus type 2 (pcv2) infection adversely affects the protective efficacy of lapinized philippines coronel (lpc) vaccine, an attenuated strain of classical swine fever virus (csfv), in pigs. in order to investigate the possible mechanisms of the pcv2-derived interference, an in vitro model was established to study the interaction of lpc virus (lpcv) and pcv2 in porcine alveolar macrophages (ams). the results showed that pcv2 reduced the lpcv infection ...201424370262
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
development of a loop-mediated isothermal amplification assay combined with a lateral flow dipstick for rapid and simple detection of classical swine fever virus in the field.classical swine fever (csf) is a highly contagious viral disease and may cause heavy economic loss to farmers. the rapid, simple and accurate diagnosis of the disease at the frontline, for example on the farms of concern is crucial for disease control. this study describes the development and evaluation of a new loop-mediated isothermal amplification (lamp) assay coupled with lateral flow dipstick (lfd) for the detection of classical swine fever virus (csfv). this rt-lamp-lfd assay combines the ...201424300833
prokaryotic expression and purification of highly soluble partial glycoprotein erns of indian strain of classical swine fever virus.classical swine fever (csf) or hog cholera, caused by a positive stranded rna virus belonging to the genus pestivirus of the flaviviridae family, is highly contagious and fatal disease of pigs. we report the novel design of construct for production of highly soluble glycoprotein erns fragment using prokaryotic expression system. a truncated fragment of the erns gene (coding for aa 109-170) denoted as 'erns-ag' was subcloned and expressed as hexa-histidine tag fusion on both terminus of protein i ...201224293833
a promising multiple-epitope recombinant vaccine against classical swine fever virus.classical swine fever (csf) is a highly contagious and often fatal disease of swine. it is caused by classical swine fever virus (csfv), one of the members of the genus pestivirus of the flaviviridae family. the development of a safe and effective vaccine against the csf is critical to pandemic control, this article shows a tandem-repeat multiple-epitope recombinant vaccine can protect pigs from csfv challenge. that was composed as following: two copies each of glycoprotein e2 residues 693-707, ...201424269058
autophagy enhances the replication of classical swine fever virus in vitro.autophagy plays an important role in cellular responses to pathogens. however, the impact of the autophagy machinery on classical swine fever virus (csfv) infection is not yet confirmed. in this study, we showed that csfv infection significantly increases the number of autophagy-like vesicles in the cytoplasm of host cells at the ultrastructural level. we also found the formation of 2 ubiquitin-like conjugation systems upon virus infection, including lc3-i/lc3-ii conversion and atg12-atg5 conjug ...201324262968
efficient generation of recombinant rna viruses using targeted recombination-mediated mutagenesis of bacterial artificial chromosomes containing full-length cdna.infectious cdna clones are a prerequisite for directed genetic manipulation of rna viruses. here, a strategy to facilitate manipulation and rescue of classical swine fever viruses (csfvs) from full-length cdnas present within bacterial artificial chromosomes (bacs) is described. this strategy allows manipulation of viral cdna by targeted recombination-mediated mutagenesis within bacteria.201324262008
innate immunity correlates with host fitness in wild boar (sus scrofa) exposed to classical swine fever.constitutive humoral immunity (chi) is thought to be a first-line of protection against pathogens invading vertebrate hosts. however, clear evidence that chi correlates with host fitness in natural conditions is still lacking. this study explores the relationship between chi, measured using a haemagglutination-haemolysis assay (hahl), and resistance to classical swine fever virus (csfv) among wild boar piglets. the individual dynamics of hahl during piglet growth was analysed, using 423 serum sa ...201324260286
the challenges of classical swine fever control: modified live and e2 subunit vaccines.classical swine fever (csf) is an economically important, highly contagious disease of swine worldwide. csf is caused by classical swine fever virus (csfv), and domestic pigs and wild boars are its only natural hosts. the two main strategies used to control csf epidemic are systematic prophylactic vaccination and a non-vaccination stamping-out policy. this review compares the protective efficacy of the routinely used modified live vaccine (mlv) and e2 subunit vaccines and summarizes the factors ...201424211665
[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
pore-forming activity of pestivirus p7 in a minimal model system supports genus-specific viroporin function.viroporins are small integral membrane proteins functional in viral assembly and egress by promoting permeabilization. blocking of viroporin function therefore constitutes a target for antiviral development. classical swine fever virus (csfv) protein p7 has been recently regarded as a class ii viroporin. here, we sought to establish the determinants of the csfv p7 permeabilizing activity in a minimal model system. assessment of an overlapping peptide library mapped the porating domain to the c-t ...201424189547
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
beta-actin interacts with the e2 protein and is involved in the early replication of classical swine fever virus.the e2 glycoprotein of classical swine fever virus (csfv) is involved in viral infection and induction of neutralizing antibodies. little is known about how e2 interacts with host cells. to understand the cellular factors involved in viral replication cycle, e2 was used as bait in yeast two-hybrid screens, resulting in the identification of β-actin as a potential e2-interacting partner. e2-β-actin interaction was confirmed by co-immunoprecipitation, gst pulldown, laser confocal and bimolecular f ...201424184320
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
a sensitive and specific nanoparticle-assisted pcr assay for rapid detection of porcine parvovirus.a novel nanoparticle-assisted polymerase chain reaction (nanopcr) assay to detect porcine parvovirus (ppv) is described here. primers for this assay were designed based on the conserved region of the nonstructural protein 1 (ns1) gene of ppv, which encodes one of the nonstructural proteins. the sensitivity of the ppv nanopcr assay was measured by using diluted recombinant plasmids in which the ppv ns1 gene had been inserted. the detection limit was 5.6 × 10(1) copies μl(-1) for the ppv nanopcr a ...201424117951
increase in chemokines cxcl10 and ccl2 in blood from pigs infected with high compared to low virulence african swine fever virus isolates.modulation of the expression of chemokines and chemokine receptors in whole blood was compared following infection of pigs with high and low virulence isolates of african swine fever virus. levels of mrnas for ccl2, ccl3l1, ccl4, cxcl10, ccr1 and ccr5 were significantly increased in at least one time point following infection in two experiments and ccl5, ccr9 and cxcr4 mrna were significantly increased in one of the experiments. the results showed that greatest fold increases in mrnas for cxcl10 ...201324083897
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
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
abortion in sows experimentally infected with african swine fever virus: clinical features.thirteen sows between 38 and 92 days of pregnancy were exposed oronasally to african swine fever virus (1979 dominican republic isolate) to determine whether infection would affect their reproductive performance. abortions occurred in sows in all stages of pregnancy. of 11 sows, 7 aborted 5 to 8 (6.1 +/- 0.90) days after inoculation or 0 to 3 (1.4 +/- 0.98) days after fever development. abortions occurred soon after clinical signs of infection first developed and viremia titers peaked. swelling ...198424049897
african swine fever virus: on the move and dangerous. 201324038277
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
autocatalytic cleavage within classical swine fever virus ns3 leads to a functional separation of protease and helicase.classical swine fever virus (csfv) is a positive-stranded rna virus belonging to the genus pestivirus within the flaviviridae family. pivotal for processing of a large portion of the viral polyprotein is a serine protease activity within nonstructural protein 3 (ns3) that also harbors helicase and ntpase activities essential for rna replication. in csfv-infected cells, ns3 appears as two forms, a fully processed ns3 of 80 kda and the precursor molecule ns2-3 of 120 kda. here we report the identi ...201323986594
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
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