prevalence of swine viral and bacterial pathogens in rodents and stray cats captured around pig farms in 2008, 102 rodents and 24 stray cats from the areas around 9 pig farms in northeast south korea were used to determine the prevalence of the following selected swine pathogens: ten viral pathogens [porcine epidemic diarrhea virus (pedv), transmissible gastroenteritis virus (tgev), rotavirus, classical swine fever virus (csfv), porcine circovirus type 2 (pcv2), encephalomyocarditis virus (emcv), porcine reproductive and respiratory syndrome virus (prrsv), porcine parvovirus (ppv), pseudorabies ...201323892461
coincidental detection of genomes of porcine parvoviruses and porcine circovirus type 2 infecting pigs in japan.the infection status of 15 viruses in 120 pigs aged about 6 months was investigated based on tonsil specimens collected from a slaughterhouse. only 5 species of porcine parvoviruses and porcine circovirus type 2 (pcv2) were detected at high frequencies; 67% for porcine parvovirus (ppv) (ppv-kr or -nadl2 as the new abbreviation), 58% for ppv2 (cnp-parv4), 39% for ppv3 (p-parv4), 33% for ppv4 (ppv4), 55% for pbo-likev (pbov7) and 80% for pcv2. a phylogenetic analysis of ppv3 suggested that japanes ...201526166811
viral inactivation of human osteochondral grafts with methylene blue and light.cartilage injury is one of the most common disorders of synovial joints. fresh osteochondral allografts are becoming a standard treatment; however, they are supply constrained with a potential risk of disease transmission. there are no known virucidal processes available for osteochondral allografts and most methods presently available are detrimental to cartilage. methylene blue light treatment has been shown to be successful in the literature for viral inactivation of fresh frozen plasma. the ...026069682
vaccine efficacy against malaria by the combination of porcine parvovirus-like particles and vaccinia virus vectors expressing cs of plasmodium.with the aim to develop an efficient and cost-effective approach to control malaria, we have generated porcine parvovirus-like particles (ppv-vlps) carrying the cd8(+) t cell epitope (syvpsaeqi) of the circumsporozoite (cs) protein from plasmodium yoelii fused to the ppv vp2 capsid protein (ppv-pycs), and tested in prime/boost protocols with poxvirus vectors for efficacy in a rodent malaria model. as a proof-of concept, we have characterized the anti-cs cd8(+) t cell response elicited by these h ...201222529915
serological prevalence of viral agents that induce reproductive failure in south korean wild boar.viral agents associated with reproductive failure such as aujeszky's disease virus (adv), encephalomyocarditis virus (emcv), and porcine parvovirus (ppv) have also been identified in european wild boar. to screen for the presence of antibodies against adv, emcv, and ppv from wild boar (sus scrofa) in south korea, 481 serum samples were collected from wild boar hunted between december 2010 and may 2011.201525888836
viral metagenomic analysis of bushpigs (potamochoerus larvatus) in uganda identifies novel variants of porcine parvovirus 4 and torque teno sus virus 1 and a result of rapidly growing human populations, intensification of livestock production and increasing exploitation of wildlife habitats for animal agriculture, the interface between wildlife, livestock and humans is expanding, with potential impacts on both domestic animal and human health. wild animals serve as reservoirs for many viruses, which may occasionally result in novel infections of domestic animals and/or the human population. given this background, we used metagenomics to investig ...201222967311
differential expression of micrornas in porcine parvovirus infected porcine cell line.porcine parvovirus (ppv), a member of the parvoviridae family, causes great economic loss in the swine industry worldwide. micrornas (mirnas) are a class of non-protein-coding genes that play many diverse and complex roles in viral infections.201526290078
complete genome sequence of porcine parvovirus 2 recovered from swine sera.a complete genomic sequence of porcine parvovirus 2 (ppv-2) was detected by viral metagenome analysis on swine sera. a phylogenetic analysis of this genome reveals that it is highly similar to previously reported north american ppv-2 genomes. the complete ppv-2 sequence is 5,426 nucleotides long.201626823583
prevalence of hiv and hcv infections in two populations of malian women and serological assays estimate the prevalence of human immunodeficiency virus (hiv) and hepatitis c virus (hcv) infections in women in mali and to evaluate the performance of serological assays.201223355914
[eukaryotic expression of ns1 major antigen region of ppv and development of an indirect elisa based on the expressed protein].to construct secretory expression vector of ppv ns1 gene, the fragment of ppv ns1 gene coding for major antigen region of the ns1 protein was amplified by pcr and inserted into multiple clone site of eukaryotic expression vector ppiczalpha-a. the recombinant ppiczalpha-a-ns1 plasmid was transferred into p. pastoris strain gs115 mediated by electro transform. recombinant p. pastoris strain gs115 was induced to express the fusion protein by methanol. the expressed and purified protein was analyzed ...201223367561
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
in vitro antiviral activity of brazilian plants (maytenus ilicifolia and aniba rosaeodora) against bovine herpesvirus type 5 and avian metapneumovirus.medicinal plants are well known for their use in traditional folk medicine as treatments for many diseases including infectious diseases.201222873798
a nanoparticle-assisted pcr assay to improve the sensitivity for rapid detection and differentiation of wild-type pseudorabies virus and gene-deleted vaccine strains.nanoparticle-assisted polymerase chain reaction (nanopcr) is a novel method for the rapid amplification of dna and has been adopted for the detection of virus because of its simplicity, rapidity, and specificity. a nanopcr assay was developed to detect and differentiate wild-type and gene-deleted pseudorabies virus (prv). three pairs of primers for nanopcr developed in this study were selected from conserved regions of prv, producing specific amplicons of 431 bp (gb), 316 bp (ge), and 202 bp (gg ...201323872268
simultaneous typing of seven porcine pathogens by multiplex pcr with a gexp analyser.a novel high-throughput method was developed for simultaneous detection and differentiation of seven porcine pathogens by multiplex pcr based on a genomelab gene expression profiler (gexp) analyser. the pathogens included in this study were pseudorabies virus (prv), classical swine fever virus (csfv), african swine fever virus (asfv), porcine reproductive and respiratory syndrome virus (prrsv), porcine parvovirus (ppv), porcine circovirus type 2 (pcv-2) and japanese encephalitis virus (jev). sev ...201626706731
biosecurity aspects and pathogen inactivation in acidified high risk animal by-products.the aim of the study was to investigate the effects of formic acid addition to ground high risk animal by-products (abp 1) in terms of stabilization and pathogen inactivation and to evaluate the biosecurity risk connected with the abp 1 based combustion fuel biomal. laboratory studies were performed on the persistence of salmonella typhimurium, bacillus cereus spores, porcine herpes virus, avian influenza virus, bovine viral diarrhea virus, equine rhinitis a virus and porcine parvovirus in bioma ...201222506709
swine infectious agents in tayassu pecari and pecari tajacu tissue samples from brazil.peccaries and pigs, tayassuidae and suidae respectively, diverged approximately one million years ago from a common ancestor. because these families share some pathogens, peccaries can act as reservoirs of infectious pathogens for domestic and wild swine. we evaluated the presence of swine infectious agents in the spleen and lung tissues of white-lipped peccaries (wlp; tayassu pecari) and collared peccaries (cp; pecari tajacu) in brazil. samples from 10 adult cp and three wlp, which had been hun ...201424484498
characterization of a novel porcine parvovirus tentatively designated ppv5.a new porcine parvovirus (ppv), provisionally designated as ppv5, was identified in u.s. pigs. cloning and sequencing from a circular or head-to-tail concatemeric array revealed that the ppv5 possesses the typical genomic organization of parvoviruses with two major predicted open reading frames (orf1 and orf2), and is most closely related to ppv4 with overall genomic identities of 64.1-67.3%. the amino acid identities between ppv5 and ppv4 were 84.6%-85.1% for orf1 and 54.0%-54.3% for orf2. unli ...201323762339
pathogen safety of a new intravenous immune globulin 10% liquid.the manufacturing process of a new intravenous immune globulin (ivig) 10% liquid product incorporates two dedicated pathogen safety steps: solvent/detergent (s/d) treatment and nanofiltration (20 nm). ion-exchange chromatography (iec) during protein purification also contributes to pathogen safety. the ability of these three process steps to inactivate/remove viruses and prions was evaluated.201728236170
inactivation of viruses during a new manufacturing process of α2-macroglobulin from cohn fraction iv by dry-heat treatment.α2-macroglobulin (α2-m) has a curative effect on radiation injury. virus transmission through plasma derivatives is still not risk-free. effect of dry heat on α2-m activity and virus inactivation by dry heat in a new manufacturing process of α2-m were studied.201627383449
evaluation of viral inactivation of pseudorabies virus, encephalomyocarditis virus, bovine viral diarrhea virus and porcine parvovirus in pancreatin of porcine origin.pancreatin is a substance containing enzymes, principally amylase, lipase, and protease. it is obtained from bovine or porcine pancreas and used in the treatment of pancreatic endocrine insufficiency in humans. regulations and safety concerns mandate viral clearance (virus removal or inactivation) in biopharmaceuticals such as pancreatin. a virus validation study was performed to evaluate virus clearance achieved in the final step of drying under vacuum by testing a panel of four animal viruses: ...201425110118
robustness of nanofiltration for increasing the viral safety margin of biological this study, the virus-removal capacity of nanofiltration was assessed using validated laboratory scale models on a wide range of viruses (pseudorabies virus; human immunodeficiency virus; bovine viral diarrhea virus; west nile virus; hepatitis a virus; murine encephalomyocarditis virus; and porcine parvovirus) with sizes from 18 nm to 200 nm and applying the different process conditions existing in a number of grifols' plasma-derived manufacturing processes (thrombin, α1-proteinase inhibitor, ...201424485384
luiii parvovirus selectively and efficiently targets, replicates in, and kills human glioma cells.because productive infection by parvoviruses requires cell division and is enhanced by oncogenic transformation, some parvoviruses may have potential utility in killing cancer cells. to identify the parvovirus(es) with the optimal oncolytic effect against human glioblastomas, we screened 12 parvoviruses at a high multiplicity of infection (moi). mvmi, mvmc, mvm-g17, tumor virus x (tvx), canine parvovirus (cpv), porcine parvovirus (ppv), rat parvovirus 1a (rpv1a), and h-3 were relatively ineffect ...201222553327
rapid and specific detection of porcine parvovirus by isothermal recombinase polymerase amplification assays.porcine parvovirus (ppv) is a major cause of swine reproductive failure and reported in many countries worldwide. recombinase polymerase amplification (rpa) assays using a real-time fluorescent detection (ppv real-time rpa assay) and a lateral flow dipstick (ppv rpa lfd assay) were developed targeting ppv ns1 gene. the detection limit of ppv real-time rpa assay was 300 copies per reaction within 9 min at 38 °c, while the rpa lfd assay has a detection limit of 400 copies per reaction in less than ...201627593155
establishment of a nanoparticle-assisted rt-pcr assay to distinguish field strains and attenuated strains of porcine epidemic diarrhea virus.porcine epidemic diarrhea virus (pedv) can cause serious disease and even death in neonatal piglets, resulting in serious damage to the swine industry worldwide. open reading frame 3 (orf3) is the only accessory gene in the pedv genome. previous studies have indicated that pedv vaccine strains have a partial deletion in orf3. in this study, a nanoparticle-assisted polymerase chain reaction (nanoparticle-assisted rt-pcr) assay targeting the orf3 of pedv was developed to distinguish pedv field str ...201627287433
simultaneous detection of eight swine reproductive and respiratory pathogens using a novel gexp analyser-based multiplex pcr assay.a new high-throughput genomelab gene expression profiler (gexp) analyser-based multiplex pcr assay was developed for the detection of eight reproductive and respiratory pathogens in swine. the reproductive and respiratory pathogens include north american porcine reproductive and respiratory syndrome virus (prrsv-na), classical swine fever virus (csfv), porcine circovirus 2 (pcv-2), swine influenza virus (siv) (including h1 and h3 subtypes), porcine parvovirus (ppv), pseudorabies virus (prv) and ...201526259690
fatal disease associated with swine hepatitis e virus and porcine circovirus 2 co-infection in four weaned pigs in recent decades, porcine circovirus 2 (pcv2) infection has been recognized as the causative agent of postweaning multisystemic wasting syndrome, and has become a threat to the swine industry. hepatitis e virus (hev) is another high prevalent pathogen in swine in many regions of the world. pcv2 and hev are both highly prevalent in pig farms in china.201525889526
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
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
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
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
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
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
development of a loop-mediated isothermal amplification assay for the detection of porcine hokovirus.hokoviruses have recently been detected as pathogens belonging to the family parvoviridae, which comprises porcine hokovirus (phov) and bovine hokovirus (bhov). in this study, we developed a loop-mediated isothermal amplification (lamp) assay for the rapid, specific and sensitive detection of phov. a set of four primers specific for six regions within the phov vp1/2 genes was designed using online software. the reaction temperature and time were optimized at 65°c and 60 min, respectively. lamp p ...201323850717
the survey of porcine teschoviruses in field samples in china with a universal rapid probe real-time rt-pcr assay.a real-time reverse transcription polymerase chain reaction (rt-pcr) based on taqman was established and evaluated for quantitative detection of porcine teschoviruses (ptvs). a pair of primers and a taqman probe targeting on the highly conserved sequence of the 5'-untranslated region (5'-utr) of one to 11 serotypes of ptv were designed. standard plasmid dna containing pcr amplification of the 5'-utr were constructed and used to develop the real-time rt-pcr. the results indicated that the real-ti ...201323138413
development of multiplex pcr for simultaneous detection of six swine dna and rna viruses.uniplex and multiplex reverse transcription-polymerase chain reaction (rt-pcr) and pcr protocols were developed and evaluated subsequently for its effectiveness in detecting simultaneously single and mixed infections in swine. specific primers for three dna viruses and three rna viruses, including classical swine fever virus (csfv), porcine reproductive and respiratory syndrome virus (prrsv), japanese encephalitis virus (jev), porcine circovirus type 2 (pcv2), porcine pseudorabies virus (prv) an ...201222575688
prevalence of and risk factors associated with viral and bacterial pathogens in farmed european wild boar.the aim of this study was to estimate in farmed european wild boars the prevalence of and risk factors associated with a range of common porcine viral and bacterial infections, namely, porcine parvovirus (ppv), porcine circovirus type 2 (pcv2), swine influenza virus (siv), aujeszky's disease virus (adv), classical swine fever virus (csfv), swine vesicular disease virus (svdv), coronavirus causing transmissible gastroenteritis (tgev), porcine reproductive and respiratory syndrome virus (prrsv), m ...201222516920
shedding patterns of endemic eurasian wild boar (sus scrofa) pathogens.the eurasianwild boar has experienced aworldwide demographic explosion that increases awareness on shared pathogens. however, shedding routes of relevant wild boar pathogens are unknown. previous observations on sex- and age-related differences in aujeszky's disease virus (adv) exposure led us to hypothesize that shedding patterns of endemicwild boar pathogens may be influenced by individual traits.we investigated shedding routes of adv, porcine parvovirus (ppv), porcine circovirus type 2 (pcv2) ...201526412545
inactivation of viruses by pasteurization at 60 °c for 10 h with and without 40% glucose as stabilizer during a new manufacturing process of α2-macroglobulin from cohn fraction iv.pasteurization is regularly used to inactivate viruses for the safety of plasma derivatives. influence of pasteurization at 60 °c for 10 h on α2-macroglobulin activity and virus inactivation were studied. with 40% sugar as stabilizers more than 70% α2-macroglobulin activity was reserved after pasteurization compared with 20% in control. glucose presented a better activity protection effect than sucrose and maltose. by pasteurization without stabilizer the virus titers of pseudorabies virus, sind ...201728215695
infectious agents identified in aborted swine fetuses in a high-density breeding area: a three-year study.reproductive failure in sows is one of the most important factors affecting pig breeding. many reproductive disorders are linked to both environmental factors and infectious agents. the goal of our study was to determine the presence of pathogens that are known to cause abortion, considering a set of conditioning factors, such as seasonality and pregnancy period. a large number of aborted fetuses (1,625 fetuses from 140 farms) from a high-density breeding area in northern italy was analyzed for ...201627400956
pathogen inactivation efficacy of mirasol prt system and intercept blood system for non-leucoreduced platelet-rich plasma-derived platelets suspended in plasma.this study was conducted to evaluate the efficacy of pathogen inactivation (pi) in non-leucoreduced platelet-rich plasma-derived platelets suspended in plasma using the mirasol prt system and the intercept blood system.201424806328
in vitro antimicrobial activities of animal-used quinoxaline 1,4-di-n-oxides against mycobacteria, mycoplasma and fungi.the quinoxaline 1,4-di-n-oxides (qdnos) were known as potent antibacterial agents. for the purpose of evaluating the bioactivity of existing animal-used qdnos drugs against representative pathogenic microorganism, the representative drugs of quinoxalines including cyadox, mequindox, quinocetone and their metabolites were submitted to the in vitro evaluation for antituberculosis, antimycoplasma, antifungal and antiviral activities.201627600955
reproduction of post-weaning multi-systemic wasting syndrome in an animal disease model as a tool for vaccine testing under controlled conditions.snatch farrowed, colostrum deprived piglets were inoculated with different combinations of porcine circovirus 2, porcine parvovirus and erysipelothrix rhusiopathiae candidate vaccines. 10 piglets were mock-vaccinated. following virus challenge with a combined porcine circovirus 2/porcine parvovirus inoculum, all animals were monitored and samples taken for serology, immunohistochemistry and qpcr. at 24 dpc all non-vaccinated animals remaining were exhibiting signs of post-weaning multi-systemic ...201627033924
health assessment of wild lowland tapir (tapirus terrestris) populations in the atlantic forest and pantanal biomes, brazil (1996-2012).abstract the lowland tapir (tapirus terrestris) is found in south america and is listed as vulnerable to extinction by the international union for conservation of nature, red list of threatened species. health issues, particularly infectious diseases, are potential threats for the species. health information from 65 wild tapirs from two brazilian biomes, atlantic forest (af) and pantanal (pa), were collected during a long-term study (1996-2012). the study included physic, hematologic and biochem ...201425105810
development of porcine rotavirus vp6 protein based elisa for differentiation of this virus and other viruses.the context and purpose of the study included 1) bacterial expression of viral protein 6 (vp6) of porcine rotavirus (prv) and generation of rabbit polyclonal antiserum to the vp6 protein; 3) establishment of a discrimination elisa to distinguish prv from a panel of other porcine viruses.201323517810
thermal treatment for pathogen inactivation as a risk mitigation strategy for safe recycling of organic waste in agriculture.thermal treatment at temperatures between 46.0°c and 55.0°c was evaluated as a method for sanitization of organic waste, a temperature interval less commonly investigated but important in connection with biological treatment processes. samples of dairy cow feces inoculated with salmonella senftenberg w775, enterococcus faecalis, bacteriophage ϕx174, and porcine parvovirus (ppv) were thermally treated using block thermostats at set temperatures in order to determine time-temperature regimes to ac ...201425035917
bioreduction of sheep carcasses effectively contains and reduces pathogen levels under operational and simulated breakdown conditions.options for the storage and disposal of animal carcasses are extremely limited in the eu after the introduction of the eu animal by-products regulations (abpr; ec/1774/2002), leading to animosity within the livestock sector and the call for alternative methods to be validated. novel storage technologies such as bioreduction may be approved under the abpr provided that they can be shown to prevent pathogen proliferation. we studied the survival of enterococcus faecalis, salmonella spp., e. coli o ...201323590844
snatch-farrowed, porcine-colostrum-deprived (sf-pcd) pigs as a model for swine infectious disease research.the current study tested the benefit of commercially available spray-dried bovine colostrum (the saskatoon colostrum company, saskatoon, saskatchewan) in raising snatch-farrowed, porcine-colostrum-deprived (sf-pcd) pigs. in experiment 1, 12 sf-pcd pigs received a liquid diet composed mainly of bovine colostrum from birth to day 10; 6 remained on the same liquid diet (col), and the other 6 were fed a diet composed mainly of milk replacer (rpl) until weaning. in experiment 2, 12 sf-pcd pigs were f ...201324082397
hepatitis e virus and porcine-derived heparin. 201323751114
inactivation effect of standard and fractionated electron beam irradiation on enveloped and non-enveloped viruses in a tendon transplant model.background: for increasing allograft tendon safety in reconstructive surgery, an effective sterilization method achieving sterility assurance including viruses without impairing the grafts properties is needed. fractionated electron beam (ebeam) has shown promising in vitro results. the proof of sufficient virus inactivation is a central part of the process validation. methods: the ebeam irradiation of the investigated viruses was performed in an optimized manner (oxygen content < 0.1%, -78 °c). ...201222896764
porcine parvovirus capsid protein expressed in escherichia coli self-assembles into virus-like particles with high immunogenicity in mice and guinea pigs.porcine parvovirus (ppv) is a causative agent of reproductive failure in pregnant sows. classical inactivated vaccine is extensively used to control ppv infection, but problems concerning safety, such as incomplete inactivation may occur. in this study, a novel subunit vaccine against ppv based on virus-like particles (vlps) formed from the complete ppv vp2 protein expressed in a prokaryotic system with co-expressed chaperones is reported. the vlps have a similar size, shape, and hemagglutinatio ...201728063996
immunogenicity of adenovirus-derived porcine parvovirus-like particles displaying b and t cell epitopes of foot-and-mouth disease.virus-like particles (vlps) vaccines combine many of the advantages of whole-virus vaccines and recombinant subunit vaccines, integrating key features that underlay their immunogenicity, safety and protective potential. we have hypothesized here the effective insertion of the vp1 epitopes (three amino acid residues 21-40, 141-160 and 200-213 in vp1, designated vpe) of foot-and-mouth disease (fmdv) within the external loops of ppv vp2 could be carried out without altering assembly based on struct ...201626685093
infectious risk factors for individual postweaning multisystemic wasting syndrome (pmws) development in pigs from affected farms in spain and denmark.two prospective longitudinal studies in 13 postweaning multisystemic wasting syndrome (pmws)-affected farms from spain (n=3) and denmark (n=10) were performed. blood samples from pigs were longitudinally collected from 1st week until the occurrence of the pmws outbreak. wasted and healthy age-matched pigs were euthanized, necropsied and histopathologically characterised. pmws diagnosis was confirmed by means of lymphoid lesions and detection of porcine circovirus type 2 (pcv2) in these tissues b ...201222884005
high-impact animal health research conducted at the usda's national animal disease center.commissioned by president dwight eisenhower in 1958 and opened with a dedication ceremony in december 1961, the usda, agricultural research service (ars), national animal disease center (nadc) celebrated its 50-year anniversary in november 2011. over these 50 years, the nadc established itself among the world's premier animal health research centers. its historic mission has been to conduct basic and applied research on selected endemic diseases of economic importance to the u.s. livestock and p ...201323642415
evaluation of a virucidal quantitative carrier test for surface disinfectants.surface disinfectants are part of broader preventive strategies preventing the transmission of bacteria, fungi and viruses in medical institutions. to evaluate their virucidal efficacy, these products must be tested with appropriate model viruses with different physico-chemical properties under conditions representing practical application in hospitals. the aim of this study was to evaluate a quantitative carrier assay. furthermore, different putative model viruses like adenovirus type 5 (adv-5) ...201424475079
antiviral effect of lithium chloride on infection of cells by porcine parvovirus.porcine parvovirus (ppv) causes reproductive failure in pigs, which leads to economic losses to the industry. as reported previously, licl efficiently impairs the replication of a variety of viruses, including the coronavirus infectious bronchitis virus (ibv), transmissible gastroenteritis virus (tgev), and pseudorabies herpesvirus. we demonstrate for the first time that inhibition of ppv replication in swine testis (st) cells by licl is dose-dependent, and that the antiviral effect of licl occu ...201525663217
a novel approach to achieving modular retrovirus clearance for a parvovirus filter.viral filtration is routinely incorporated into the downstream purification processes for the production of biologics produced in mammalian cell cultures (mcc) to remove potential viral contaminants. in recent years, the use of retentive filters designed for retaining parvovirus (~20 nm) has become an industry standard in a conscious effort to further improve product safety. since retentive filters remove viruses primarily by the size exclusion mechanism, it is expected that filters designed for ...201424123923
mechanistic evaluation of virus clearance by depth filtration.virus clearance by depth filtration has not been well-understood mechanistically due to lack of quantitative data on filter charge characteristics and absence of systematic studies. it is generally believed that both electrostatic interactions and sized based mechanical entrapment contribute to virus clearance by depth filtration. in order to establish whether the effectiveness of virus clearance correlates with the charge characteristics of a given depth filter, a counter-ion displacement techn ...201525683459
identification of mycoplasma suis antigens and development of a multiplex microbead immunoassay.the aims of the current study were to identify mycoplasma suis antigens and develop a multiplex microbead immunoassay (mia). a m. suis-expression library was screened for immunogens using sera from infected pigs. based on bioinformatics, putative antigens were identified within positive inserts; gene fragments were expressed and purified as polyhistidine fusion proteins, and immunoreactivity was confirmed by western blot. selected antigens were used to develop a mia. sera from noninfected and in ...201424557752
cd8(+) t-cell-mediated immunity against malaria: a novel heterologous prime-boost strategy.recently, a vaccine against malaria was successfully tested in a human phase iii trial. the efficacy of this vaccine formulation, based on the plasmodium falciparum circumsporozoite protein, was approximately 50% and correlated with the presence of antibodies specific to the infective stages of the malaria parasites. different strategies are being pursued to improve vaccine efficacy levels. one such strategy is the induction of specific cytotoxic t cells that can destroy the intracellular hepato ...201223151162
reduction of spiked porcine circovirus during the manufacture of a vero cell-derived vaccine.porcine circovirus-1 (pcv1) was recently identified as a contaminant in live rotavirus vaccines, which was likely caused by contaminated porcine trypsin. the event triggered the development of new regulatory guidance on the use of porcine trypsin which shall ensure that cell lines and porcine trypsin in use are free from pcv1. in addition, manufacturing processes of biologicals other than live vaccines include virus clearance steps that may prevent and mitigate any potential virus contamination ...201424560672
a comprehensive microbiological safety approach for agarose encapsulated porcine islets intended for clinical trials.the use of porcine islets to replace insulin-producing islet β-cells, destroyed during the diabetogenic disease process, presents distinct challenges if this option is to become a therapeutic reality for the treatment of type 1 diabetes. these challenges include a thorough evaluation of the microbiological safety of the islets. in this study, we describe a robust porcine islet-screening program that provides a high level of confidence in the microbiological safety of porcine islets suitable for ...201627862363
experimental and computational surface hydrophobicity analysis of a non-enveloped virus and proteins.the physical characteristics of viruses needs to be understood in order to manipulate the interaction of viruses with host cells, as well as to create specific molecular recognition techniques to detect, purify, and remove viruses. viruses are generally believed to be positively charged at physiological ph, but there are few other defining characteristics. here, we have experimentally and computationally demonstrated that a non-enveloped virus is more hydrophobic than a panel of model proteins. ...201728219841
epidemiological and immunopathological studies on porcine parvovirus infection in punjab.the aim of this study was to get the first-hand knowledge about the seroprevalence of porcine parvovirus (ppv) in punjab and a diagnosis of ppv from abortion cases of swine using gross, histopathological, and immunohistopathological techniques to observe the tissue tropism of the virus strain.201627651669
[immune response of recombinant pseudorabies virus rprv-vp2 expressing vp2 gene of porcine parvovirus in mice].in order to develop a combined live vaccine that will be used to prevent against porcine parvovirus (ppv) and pseudorabies virus (prv) infection, the vp2 gene of ppv was inserted into the transfer vector plasmid pg to produce the recombinant plasmid pgvp2. the plasmid pgvp2 and the genome of prv hb98 attenuated vaccine were transfected by using lipofectamine into swine testis cells for the homologous recombination. the recombinant virus rprv-vp2 was purified by selection of green fluorescence pl ...201627396164
removal process of prion and parvovirus from human platelet lysates used as clinical-grade supplement for ex vivo cell expansion.pooled human platelet lysate (hpl) is becoming the new gold standard as supplement for ex vivo cell culture for clinical protocols. however, the risk of pathogen contamination of hpl increases with the platelet pool size. we hypothesized that hollow fiber anion exchange membrane chromatography using qyuspeed d (qsd) could remove resistant and untested bloodborne pathogens, such as parvoviruses and prions, from hpl-supplemented growth media without substantially affecting their capacity to suppor ...201627260210
an inactivated whole-virus porcine parvovirus vaccine protects pigs against disease but does not prevent virus shedding even after homologous virus challenge.inactivated whole-virus vaccines against porcine parvovirus (ppv) can prevent disease but not infection and virus shedding after heterologous virus challenge. here, we showed that the same is true for a homologous challenge. pregnant sows were vaccinated with an experimental inactivated vaccine based on ppv strain 27a. they were challenged on day 40 of gestation with the virulent porcine parvovirus ppv-27a from which the vaccine was prepared (homologous challenge). on day 90 of gestation, the fe ...201626939976
porcine parvovirus as a contaminant in cell cultures and laboratory supplies.although ppv has been described as a cellular contaminant, few recent studies about the presence of this virus in cell cultures, serum, and trypsin were found in the literature. the purpose of this study was to detect the presence of porcine parvovirus (ppv) by polymerase chain reaction (pcr) in cell cultures, serum, and trypsin used in official public laboratories of educational institutes and research centers. we tested samples of cell cultures (88), batches of trypsin (10), and fetal bovine s ...201626811218
first identification of porcine parvovirus 6 in north america by viral metagenomic sequencing of serum from pigs infected with porcine reproductive and respiratory syndrome virus.currently, eight species in four genera of parvovirus have been described that infect swine. these include ungulate protoparvovirus 1 (classical porcine parvovirus, ppv), ungulate tetraparvovirus 2 (ppv3), ungulate tetraparvovirus 3 (which includes ppv2, porcine hokovirus, porcine partetravirus and porcine parv4), ungulate copiparvovirus 2 (which includes ppv4 and ppv5), ungulate bocaparvovirus 2 (which includes porcine bocavirus 1, 2 and 6), ungulate bocaparvovirus 3 (porcine bocavirus 5), ungu ...201526475593
molecular epidemiology and evolution of porcine parvoviruses.porcine parvovirus (ppv), recently named ungulate protoparvovirus 1, is considered to be one of the most important causes of reproductive failure in swine. fetal death, mummification, stillbirths and delayed return to estrus are predominant clinical signs commonly associated with ppv infection in a herd. it has recently been shown that certain parvoviruses exhibit a nucleotide substitution rate close to that commonly determined for rna viruses. however, the ppv vaccines broadly used in the last ...201526453771
the pathogenic role of torque teno sus virus 1 and 2 and their correlations with various viral pathogens and host immunocytes in wasting pigs.the pathogenic role of torque teno sus virus (ttsuv) in swine is controversial among different studies. the present study intended to evaluate the potential pathogenicity of ttsuv based on its correlations with the histopathological changes, various common concurrently infected viral pathogens including porcine circovirus type 2 (pcv2), porcine reproductive and respiratory syndrome virus (prrsv), and porcine parvovirus (ppv), as well as changes in the distribution and population of host immunocy ...201526390821
enveloped virus flocculation and removal in osmolyte solutions.our ability to reduce infectious disease burden throughout the world has been greatly improved by the creation of vaccines. however, worldwide immunization rates are low. the two most likely reasons are the lack of sufficient distribution in underdeveloped countries and the high cost of vaccine products. the high costs are due to the difficulties of manufacturing individual vaccine products with specialized purification trains. in this study, we propose to use virus flocculation in osmolytes, fo ...201525865274
in vitro antiviral activity of germacrone against porcine parvovirus.porcine parvovirus (ppv) infections can lead to significant losses to the swine industry by causing reproductive failure in pigs. germacrone has been reported to efficiently suppress the replication of influenza virus. in this report, the antiviral activity of germacrone on ppv in swine testis (st) cells was investigated. here, we show for the first time that germacrone protects cells from ppv infection and suppresses the synthesis of viral mrna and protein. furthermore, we show that germacrone ...201525813663
separation of porcine parvovirus from bovine serum albumin using peg-salt aqueous two-phase system.vaccine production faces a challenge in adopting conventional downstream processing steps that can efficiently purify large viral particles. some major issues that plague vaccine purification are purity, potency, and quality. the industry currently considers 30% as an acceptable virus recovery for a vaccine purification process, including all downstream processes, whereas antibody recovery from cho cell culture is generally around 80-85%. a platform technology with an improved virus recovery wou ...201425086421
classic nuclear localization signals and a novel nuclear localization motif are required for nuclear transport of porcine parvovirus capsid proteins.nuclear targeting of capsid proteins (vps) is important for genome delivery and precedes assembly in the replication cycle of porcine parvovirus (ppv). clusters of basic amino acids, corresponding to potential nuclear localization signals (nls), were found only in the unique region of vp1 (vp1up, for vp1 unique part). of the five identified basic regions (br), three were important for nuclear localization of vp1up: br1 was a classic pat7 nls, and the combination of br4 and br5 was a classic bipa ...201425078698
generation of recombinant porcine parvovirus virus-like particles in saccharomyces cerevisiae and development of virus-specific monoclonal antibodies.porcine parvovirus (ppv) is a widespread infectious virus that causes serious reproductive diseases of swine and death of piglets. the gene coding for the major capsid protein vp2 of ppv was amplified using viral nucleic acid extract from swine serum and inserted into yeast saccharomyces cerevisiae expression plasmid. recombinant ppv vp2 protein was efficiently expressed in yeast and purified using density gradient centrifugation. electron microscopy analysis of purified ppv vp2 protein revealed ...201425045718
porcine parvovirus flocculation and removal in the presence of osmolytes.viruses can be modified into viral vaccines or gene therapy vectors in order to treat acquired or genetic diseases. to satisfy the current market demand, an improvement in current vaccine manufacturing is needed. chromatography and nanofiltration are not suitable for all types of viruses. in this study, we propose to use virus flocculation with osmolytes, followed by microfiltration, as a potential virus purification process. we hypothesize that osmolytes strongly bind to water, thus leading to ...201425003646
adsorption of a non-enveloped mammalian virus to functionalized the pursuit of finding superior methods to remove pathogens from drinking water, this study examines the adsorption of a non-enveloped, mammalian virus to highly charged nanofibers. n-[(2-hydroxyl-3-trimethylammonium) propyl] chitosan (htcc) nanofibers were synthesized by the addition of a quaternary amine to chitosan. htcc was blended with polyvinyl alcohol (pva) to produce nanofibers by electrospinning. the nanofibers were stabilized against water by crosslinking with glutaraldehyde. when s ...201424985761
antiviral effect of diammonium glycyrrhizinate on cell infection by porcine parvovirus.porcine parvovirus (ppv) can cause reproductive failure in swine, resulting in economic losses to the industry. antiviral effects of diammonium glycyrrhizinate (dg) have been reported on several animal viruses; however, to date it has yet to be tested on ppv. in this study, the antiviral activity of dg on swine testis (st) cell infection by ppv was investigated using an empirically determined, non-toxic concentration of dg and three different experimental designs: (1) pre-treatment of virus prio ...201424614970
differential replication of two porcine parvovirus strains in bovine cell lines ensues from initial dna processing and ns1 expression.porcine parvovirus (ppv) is a small dna virus with restricted coding capacity. the 5 kb genome expresses three major non-structural proteins (ns1, ns2 and sat), and two structural proteins (vp1 and vp2). these few viral proteins are pleiotropic and interact with cellular components throughout viral replication. in this regard, very few cell lines have been shown to replicate the virus efficiently. cell lines were established from a primary culture of bovine cells that allowed allotropic variants ...201424347493
non-enveloped virus reduction with quaternized chitosan nanofibers containing graphene.membranes are an accepted technology for water purification. membrane filtration can remove pathogens, including bacteria and viruses, by size. for small viruses that can have a diameter <25nm, removal by size leads to large membrane areas, high transmembrane pressures, low water flux, and frequent changing of membranes. in this work, we discovered that electrospun nanofibers made of chitosan and functionalized with a quaternary amine (htcc) have the ability to adsorb a model non-enveloped virus ...201324041631
[proliferation characteristics of a pk-15 cell-adapted strain of porcine parvovirus].to study the proliferation characteristics of ppv in differently infected way and the variance of concentrations in different cells. a strain of porcine parvovirus(ppv) was adapted to pk-15 cells, and a real-time fluorescent quantitative pcr (fq-pcr) assay was developed based on the specific region of the ns1 gene of ppv to quantify the ppv. the fq-pcr was used to measure the viral concentration of virus-infected cells by simultaneous or step by step inoculation and plot one-step growth curves. ...201323894997
population dynamics and in vitro antibody pressure of porcine parvovirus indicate a decrease in estimate the impact of porcine parvovirus (ppv) vaccines on the emergence of new phenotypes, the population dynamic history of the virus was calculated using the bayesian markov chain monte carlo method with a bayesian skyline coalescent model. additionally, an in vitro model was performed with consecutive passages of the 'challenge' strain (a virulent field strain) and nadl2 strain (a vaccine strain) in a pk-15 cell line supplemented with polyclonal antibodies raised against the vaccine stra ...201323804570
reduction of porcine parvovirus infectivity in the presence of protecting osmolytes.osmolytes are natural compounds found in the cells of many organisms that stabilize intracellular proteins against environmental stresses. protecting osmolytes can promote protein folding, whereas denaturing osmolytes have the opposite effect. a variety of osmolytes were tested for their antiviral activity against porcine parvovirus (ppv). ppv is a non-enveloped, icosahedral, single-strand dna virus. we have discovered two protecting osmolytes, trimethylamine n-oxide (tmao) and glycine that redu ...201323648707
influence of minor displacements in loops of the porcine parvovirus vp2 capsid on virus-like particles assembly and the induction of antibody antigen-delivery system based on hybrid virus-like particles (vlps) formed by the self-assembly of the capsid vp2 protein of porcine parvovirus (ppv) and expressing foreign peptides offers an alternative method for vaccination. in this study, the three-dimensional structure of the ppv capsid protein and surface loops deletion mutants were analyzed to define essential domains in ppv vp2 for the assembly of vlps. electron microscopic analysis and sds-page analysis confirmed the presence of abun ...201323430711
complete genome sequence of a novel porcine parvovirus (ppv) provisionally designated ppv5.a new porcine parvovirus (ppv) was identified from pig lung tissues in the united states. it is most closely related to the recently identified ppv4 with overall genomic identities of 64.1 to 67.3%. unlike ppv4, this virus lacks the additional open reading frame 3 (orf3) and has a much longer orf2. the name ppv5 is provisionally proposed.201323405295
interclade recombination in porcine parvovirus strains.a detailed analysis of the ns1/vp1vp2 genome region of the porcine parvovirus (ppv) strains isolated from vaccinated animals was performed. we found many inconsistencies in the phylogenetic trees of these viral isolates, such as low statistical support and strains with long branches in the phylogenetic trees. thus, we used distance-based and phylogenetic methods to distinguish de facto recombinants from spurious recombination signals. we found a mosaic virus in which the ns1 gene was acquired fr ...201222993191
serum protein profiles as potential biomarkers for infectious disease status in veterinary medicine and animal husbandry, there is a need for tools allowing the early warning of diseases. preferably, tests should be available that warn farmers and veterinarians during the incubation periods of disease and before the onset of clinical signs. the objective of this study was to explore the potential of serum protein profiles as an early biomarker for infectious disease status. serum samples were obtained from an experimental pig model for porcine circovirus-associated disea ...201222439879
susceptibility of porcine preimplantation embryos to viruses associated with reproductive the modern biological area, the applications of pig as a laboratory model have extensive prospects, such as gene transfer, ivf, scnt, and xenotransplantation. however, the risk of pathogen transmission by porcine embryos is always a topic to be investigated, especially the viruses related to reproductive failure, for instance, pseudorabies virus, porcine reproductive and respiratory syndrome virus, porcine parvovirus, and porcine circovirus type 2. it should be mentioned that the zona pelluci ...201627423729
development of real-time pcr assay for detection of porcine circovirus-like virus p1 in domestic pigs in china.the porcine circovirus-like agent p1 is a newly discovered dna virus with a single-stranded circular genome that is highly homologous to that of porcine circovirus type 2. p1 infection can cause symptoms resembling postweaning multisystemic wasting syndrome. this study aims to develop a rapid, sensitive and specific method to detect p1.201526404908
characterization of porcine parvovirus type 3 and porcine circovirus type 2 in wild boars (sus scrofa) in the number of free-living wild boars (sus scrofa l.) continues to rise in slovakia, the probability of pathogen transmission between susceptible species increases. we investigated the distribution and genetic characterization of porcine parvovirus type 3 (ppv3), porcine circovirus type 2 (pcv2), and their coinfection in wild boars. among 194 animals tested, 19.1% were positive for ppv3 and 43.8% for pcv2. similar rates of coinfection with both viruses reaching 11.0% and 11.8% were observed in ...201525973618
a taqman-based real-time pcr for detection and quantification of porcine parvovirus 4.porcine parvovirus 4 (ppv4) is a dna virus, and a member of the parvoviridae family within the bocavirus genera. it was detected recently in swine, but its epidemiology and pathology remain unclear. a taqman-based real-time pcr (qpcr) targeting a conserved region of the orf3 gene of ppv4 was developed. the qpcr detection limit was 9.5 × 10(1) dna copies/μl. there was no cross-reaction with porcine parvovirus, torque teno virus 1, torque teno virus 2, porcine circovirus type 1, porcine circovirus ...201525813599
investigating risk factors and possible infectious aetiologies of mummified fetuses on a large piggery in investigate risk factors and potential infectious aetiologies of an increased mummification rate (>2%) identified over time on a 1200-sow farrow-to-finish farm in australia.201425424759
identification of recently described porcine parvoviruses in archived north american samples from 1996 and association with porcine circovirus associated disease.the association of porcine circovirus (pcv) type 2 and porcine parvovirus (ppv) type 1 as a cause of porcine circovirus associated disease (pcvad) is well established. the objective of this study was to investigate the prevalence rates of classical ppv1 and recently recognized ppv2-5 in serum and lung samples from pigs and farms with known pcv2 status. a total of 586 serum samples and 164 lung homogenates collected from 1996 to 2013 in the usa and canada were utilized. all samples were tested fo ...201425081955
development and evaluation of a loop-mediated isothermal amplification method for rapid detection and differentiation of two genotypes of porcine circovirus type 2.porcine circovirus type 2 (pcv2) is one of the major swine viral diseases and has caused significant economic loss to pig producers. pcv2 has been divided into two major genotypes: pcv2a, pcv2b. a loop-mediated isothermal amplification (lamp) method was developed for the detection and differentiation of pcv2a and pcv2b in clinical samples.201423845855
a pcv2 vaccine based on genotype 2b is more effective than a 2a-based vaccine to protect against pcv2b or combined pcv2a/2b viremia in pigs with concurrent pcv2, prrsv and ppv infection.the predominant genotype of porcine circovirus (pcv) in the pig population today is pcv2b yet pcv2a-based commercial vaccines are considered effective in protecting against porcine circovirus associated disease. the objective of this study was to compare the ability of pcv2a- and pcv2b-based vaccines to control pcv2b viremia in a challenge model that mimics the u.s. field situation. sixty-three pigs were randomly assigned to one of eight groups. sixteen pigs were vaccinated with an experimental ...201323174198
long-term monitoring of 10 selected pathogens in wild boar (sus scrofa) in sierra nevada national park, southern spain.wild boar (sus scrofa) populations are increasing in the iberian peninsula, and population management must include disease management and control. in this study, the epidemiology of 10 selected pathogens (aujeszky's disease virus - adv, porcine reproductive and respiratory syndrome virus - prrsv, porcine influenza virus, porcine circovirus, porcine parvovirus, erysipelotrix rhusiopathiae, leptospira pomona, chlamydia/chlamydiaceae sp., salmonella sp. and mycobacterium bovis) in the wild boar pop ...201425261921
lack of antibody protection against porcine circovirus 2 and porcine parvovirus in naturally infected dams and their offspring.the aim of this study was to evaluate porcine parvovirus (ppv) and porcine circovirus 2 (pcv2) infection in dams and their offspring and the role of antibody protection on these infections. sera were collected from gilts and sows by venipuncture and from umbilical cord of newborn pre-suckle piglets for the detection of pcv2 and ppv antibodies by immunoperoxidase monolayer and haemmaglutination inhibition assays, respectively. gilts and sows sera were submitted to viral detection by pcr, as well ...201323084257
[infection related fertility disorders in swiss pig breeding farms at the end of the postweaning multisystemic wasting syndrome (pmws) epizooty].porcine circovirus type 2 (pcv2) is able to induce reproductive failures. 286 fetuses from 113 sows of 59 farms with increased reproductive disorders which included abortions, mummies, stillborn and weak born piglets were studied six years after the beginning of the epizooty of postweaning multisystemic wasting syndrome (pmws) in switzerland. 14 % of the cases were bacterial infections based on histological signs of inflammation and pathogen isolation. 12 % further cases showed inflammatory reac ...201223027510
simultaneous detection of porcine parvovirus and porcine circovirus type 2 by duplex real-time pcr and amplicon melting curve analysis using sybr green.the development of a sybr green-based duplex real-time pcr is described for simultaneous detection of porcine parvovirus (ppv) and porcine circovirus type 2 (pcv-2) genomes. viral genomes were identified in the same sample by their distinctive melting temperature (t(m)) which is 77.5°c for ppv vp2 313bp amplicon and 82.3°c for pcv-2 orf2 171bp amplicon, respectively. the detection limit of the method was 0.01tcid(50)/ml for ppv and pcv-2, about 10 times more sensitive than conventional pcr. in a ...201322771739
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