a viral phospholipase a2 is required for parvovirus infectivity.sequence analysis revealed phospholipase a2 (pla2) motifs in capsid proteins of parvoviruses. although pla2 activity is not known to exist in viruses, putative pla2s from divergent parvoviruses, human b19, porcine parvovirus, and insect gmdnv (densovirus from galleria mellonella), can emulate catalytic properties of secreted pla2. mutations of critical amino acids strongly reduce both pla2 activity and, proportionally, viral infectivity, but cell surface attachment, entry, and endocytosis by pla ...200111702787
the structure of porcine parvovirus: comparison with related viruses.the structure of baculovirus-expressed porcine parvovirus (ppv) capsids was solved using x-ray crystallography and was found to be similar to the related canine parvovirus (cpv) and minute virus of mice (mvm). the ppv capsid protein has 57 % and 49 % amino acid sequence identity with cpv and mvm, respectively, but the degree of conservation of surface-exposed residues is lower than average. consequently, most of the structural differences are on the surface and are the probable cause of the know ...200211827486
virus safety of a porcine-derived medical device: evaluation of a viral inactivation method.the goal of this study was to evaluate the efficacy of a virus-inactivating process for use during the preparation of porcine-derived extracellular matrix biomaterials for human clinical implantation. porcine small intestine, the source material for the tissue-engineered, small intestinal submucosa (sis) biomaterial, was evaluated. relevant enveloped, non-enveloped, and model viruses representative of different virus families were included in the investigation: porcine parvovirus (ppv), porcine ...200212115437
the capsid protein of a plant single-stranded rna virus is modified by o-linked n-acetylglucosamine.plum pox virus (ppv) is a member of the potyvirus genus of plant viruses. labeling with udp-[3h]galactose and galactosyltransferase indicated that the capsid protein (cp) of ppv is a glycoprotein with n-acetylglucosamine terminal residues. mass spectrometry analysis of different ppv isolates and mutants revealed o-linked n-acetylglucosamination, a modification barely studied in plant proteins, of serine and/or threonine residues near the amino end of ppv cp. cp of ppv virions is also modified by ...200211606576
clinical and economic implications of adding hpv tests to the routine cytology follow-up and management of patients with histologically defined cervical intraepithelial neoplasia grade 1.the objective of this study was to evaluate the clinical and economic implications of adding human papillomavirus (hpv) testing to the follow-up and management protocol of women with a histological diagnosis of low-grade cervical intraepithelial neoplasia (cin1).200212144817
insect cell culture medium supplementation with fetal bovine serum and bovine serum albumin: effects on baculovirus adsorption and infection kinetics.the effects of insect cell culture medium supplementation with fbs were investigated. bsa was found to be the factor responsible for the increased baculovirus infection rate of fbs-supplemented cultures in a concentration-dependent form up to 25 g l(-)(1). lower rates of baculovirus binding to cells were observed with fbs- and bsa-supplemented cultures compared with infections carried out in serum-free media. virus attachment constants were found to depend on medium matrix composition. an effici ...200212153321
predictive value of alt levels for histologic findings in chronic hepatitis c: a european collaborative study.the aim of this retrospective study was to determine the predictive value of alanine aminotransferase (alt) levels for histologic findings in patients with chronic hepatitis c virus (hcv) infection. data on 864 hcv rna-positive patients were collected. alt values were obtained at the time of biopsy (before treatment), and normal alt values were defined as normal values obtained at serial evaluations during a 6-month period. histologic results were scored using the metavir system. among all patie ...200212297846
endemic hepatitis c virus infection in a sicilian town: further evidence for iatrogenic transmission.the prevalence of and risk factors for hcv and hbv infections in the general population and the predictive value of alt screening in identifying anti-hcv positive subjects have been evaluated in a small sicilian town. a random 1:4 sampling from the census of the general population was performed. anti-hcv, hcv-rna, hcv genotype, hbsag, and anti-hbc were tested. the linkage between hcv infection and potential risk factors was evaluated by multiple logistic regression analysis. among 721 subjects s ...200212116024
integrated process optimization: lessons from retrovirus and virus-like particle production.the optimization of production and purification processes is usually approached by engineers from a strictly biotechnological point of view. the present paper envisages the definition and application of an optimization model that takes into account the impact of both biological and technological issues upon the optimization protocols and strategies. for this purpose, the optimization of three analogous but different systems comprising animal cell growth and bioparticle production is presented. t ...200212385709
the production of a genus-specific recombinant antibody (scfv) using a recombinant potyvirus protease.a single chain variable fragment antibody (scfv; anti-nia scfv102) was selected from a synthetic human antibody library by using a nia protease of plum pox virus (ppv) as an antigen, which was expressed in bacteria. the nia protease forms the nuclear inclusion body a and acts as the major protease in the cleavage of the viral polyprotein into functional proteins. the nia protein was detected with anti-nia scfv102 after expression in escherichia coli cells as well as from ppv-infected nicotiana b ...200212393153
the evidence behind inhibitor treatment with porcine factor viii.factor viii auto- and alloantibodies neutralise porcine factor viii to a lesser extent than factor viii of human origin. the reduced reactivity of the porcine molecule, predominantly due to sequence variation in the a2 and c2 domains, has been the rationale for using porcine factor viii to secure haemostasis for patients with factor viii inhibitors. porcine factor viii has been shown to provide effective haemostatic control particularly for patients with intermediate inhibitor titres with limite ...200212214138
case-control study on the association of porcine circovirus type 2 and other swine viral pathogens with postweaning multisystemic wasting syndrome.a field-based case-control study was conducted to assess the strength of association of porcine circovirus type 2 (pcv2) and some major swine viruses with postweaning multisystemic wasting syndrome (pmws). cases were defined as individual pigs with a clinical history of progressive weight loss and histopathological lesions characteristic of pmws. controls were pigs without clinical signs and histopathological lesions typical of pmws. a total of 31 cases and 56 controls was identified from diagno ...200212423025
the etiology and management of genital ulcers in the dominican republic and peru.clinical diagnosis of genital ulcers is difficult, and diagnostic tests are least available in settings where rates of disease are highest. the world health organization (who) has developed protocols for the syndromic management of genital ulcers in resource-poor settings. however, because risk factors, patterns and causes of disease, and antimicrobial susceptibilities differ from region to region and over time, they must be adapted to local situations.200212370522
pathogen inactivation of blood components: current status and introduction of an approach using riboflavin as a photosensitizer.riboflavin is a naturally occurring compound and an essential human nutrient. studies in the 1960s and 70s showed that it could be effective, when exposed to visible or uv light, in inactivating viruses and bacteria. this suggested to us that it could act as a photosensitizer useful in the inactivation of pathogens found in blood products, because of its nucleic acid specificity and its limited tendency toward indiscriminate oxidation. the riboflavin molecule is a planar, conjugated ring structu ...200212430933
broad-spectrum virus reduction in red cell concentrates using inactine pen110 chemistry.the risk of transmission of blood-borne pathogens by transfusion is a persistent problem in medicine. to address this safety issue, inactine pen110 chemistry is being utilized to develop a process for preparing pathogen-reduced red blood cell concentrates (rbcc). the purpose of this study was to characterize the virucidal effectiveness of the inactine pen110 chemistry in full units of rbcc by using a panel of viruses with diverse properties in composition, size and shape.200212437518
effective use of gamma irradiation for pathogen inactivation of monoclonal antibody preparations.gamma irradiation has been used for decades as an effective method of pathogen inactivation of relatively inert materials. until recently, its application to biologicals has resulted in unacceptable losses in functional activity. in this report we demonstrate that the damaging secondary effects of gamma irradiation can be controlled while maintaining the pathogen inactivation properties due to damage by primary effects. control is achieved by a combination of protection from free radical damage ...200212217345
prevalence of porcine reproductive and respiratory syndrome virus, porcine circovirus type 2 and porcine parvovirus from aborted fetuses and pigs with respiratory problems in korea.porcine reproductive and respiratory syndrome virus(prrsv)0, porcine circovirus type 2(pcv-2) and porcine parvovirus (ppv)0 infections were investigated as possible causes of the postweaning multisystemic wasting syndrome(pmws). specific primers for rt-pcr and pcr were designed for the differential detection of prrsv, pcv-2 and ppv. using pcr, these viruses were detected in homogenized tissue samples from pigs that had respiratory of reproductive problems in the time period between 1998 and 2000 ...200112441689
viral pharmacovigilance study of haemophiliacs receiving porcine factor viii.porcine factor viii (fviii; hyate:c; speywood biopharm ltd, uk) has been used since 1980 for the treatment both of patients with acquired haemophilia and those with congenital haemophilia and inhibitory antibodies. each batch is extensively screened using cell-culture techniques to confirm the absence of viruses. the production process does not incorporate specific virucidal treatment steps, such as heat treatment or the addition of a solvent/detergent mixture. low levels of porcine parvovirus w ...200212410650
usefulness of dna ploidy measurement on liquid-based smears showing conflicting results between cytology and high-risk human papillomavirus improve the positive predictive value (ppv) for high-risk human papillomavirus (hr-hpv) in primary screening, dna ploidy was measured on the same liquid-based sample by image cytometry in 984 cases showing discrepancies between cytology and hr-hpv testing. of the conflicting results, 14.5% corresponded to a cytologic lesion (from atypical squamous cells of undetermined significance to high-grade squamous intraepithelial lesion [hsil]) without hpv detected, and 85.5% of smears were within norm ...200212428790
antibodies to selected viral and bacterial pathogens in european wild boars from southcentral spain.serum samples from 78 european wild boars (sus scrofa) harvested during the 1999-2000 hunting season were tested for antibodies to brucella spp., classical swine fever virus, erysipelothrix rhusiopathiae, haemophilus parasuis, leptospira interrogans serovar pomona, mycoplasma hyopneumoniae, pseudorabies virus (prv), porcine parvovirus (ppv), porcine reproductive and respiratory syndrome virus, salmonella serogroups b, c, and d, streptococcus suis, and swine influenza virus (siv) serotypes h1n1 a ...200212238391
identification of immunogenic hot spots within plum pox potyvirus capsid protein for efficient antigen presentation.pepscan analysis has been used to characterize the immunogenic regions of the capsid protein (cp) in virions of plum pox potyvirus (ppv). in addition to the well-known highly immunogenic n- and c-terminal domains of cp, regions within the core domain of the protein have also shown high immunogenicity. moreover, the n terminus of cp is not homogeneously immunogenic, alternatively showing regions frequently recognized by antibodies and others that are not recognized at all. these results have help ...200212438590
incorporation of an additional viral-clearance step into a human immunoglobulin manufacturing process.regulatory agencies have mandated that manufacturers of immunoglobulin products incorporate robust viral inactivation or removal steps into their purification processes. we evaluated the effectiveness of incorporating nanofiltration, a generic virus-clearance step, into an existing plasma-fractionation process for a human anti-d immunoglobulin product.200212366764
generation and characterisation of functional recombinant antibody fragments against rna replicase nib from plum pox virus.a monoclonal antibody (mab 2a) able to react against the rna replicase nib from plum pox virus (ppv) was obtained and used for generating a specific scfv fragment. the vh and vl coding sequences were cloned and expressed as a fusion scfv protein to alkaline phosphatase. this fusion protein was able to recognise viral nib in both western and tissue-print elisa blots. the affinity and specificity of scfv2a for nib was similar to that of the parental mab and the region yleafy from ppv-nib was ident ...200312535657
inactivation of viral and prion pathogens by gamma-irradiation under conditions that maintain the integrity of human albumin.the administration of therapeutic plasma protein concentrates has been associated with the real risk of transmitting viral diseases and the theoretical risks of prion transmission. our objective was to determine if gamma-irradiation can inactivate viral or prion infectivity without damaging a protein biotherapeutically.200312542732
immunologic features of porcine circovirus type 2 infection.clinical expression of porcine circovirus 2 (pcv-2) infection in swine may result in two distinct high mortality disease syndromes. in north america, postweaning multisystemic wasting syndrome (pmws), while still sporadic in incidence, predominates. in europe and elsewhere, both pmws and a second syndrome, porcine dermatitis and nephropathy syndrome (pdns), occur in endemic and epidemic forms. pmws but not pdns has been reproduced in piglets by inoculations with pcv-2 alone or in pcv-2-infected ...200212513928
molecular studies of the synergistic interactions between plum pox virus hc-pro protein and potato virus x.helper component proteinase (hc-pro) is a multifunctional viral protein involved in vection and movement of potyvirus; suppression of host post-transcriptional gene silencing reaction; and synergism of potyvirus with other viruses, notably potexvirus. when the hc-pro of plum pox potyvirus (ppv) was transiently expressed in n. benthamiana and n. clevelandii plants via potato virus x (pvx) vector, a highly synergistic response of leaf necrosis or plant death, was observed in pvxhc-infected plants. ...200212491099
inactivation of parvovirus b19 during pasteurization of human serum has been shown that hsa may be contaminated with parvovirus b19 (b19) dna. however, the presence of b19 dna does not necessarily indicate infectious virus. hsa is pasteurized at 60 degrees c for 10 hours and it remains unclear whether this procedure inactivates b19. studies with animal parvoviruses indicate considerable heat resistance at 60 degrees c. however, due to the lack of a suitable cell culture system, the pasteurization process has not been investigated in the past.200212385412
comparison of the denka-seiken influ a.b-quick and bd directigen flu a+b kits with direct fluorescent-antibody staining and shell vial culture methods for rapid detection of influenza viruses.the influ a.b-quick and directigen flu a+b enzyme immunoassays were compared with direct immunofluorescence and cell culture for detection of influenza a and b viruses in a total of 255 patient specimens. both assays identified 23 of 42 influenza a viruses (sensitivity, 54.8%; specificity, 100%; positive predictive value [ppv], 100%; negative predictive value [npv], 91.8%). the influ a.b-quick assay identified 10 of 16 influenza b viruses (sensitivity, 62.5%; specificity, 99.6%; ppv, 90.9%; npv, ...200312734274
comparative serologic and virologic study of commercial swine herds with and without postweaning multisystemic wasting syndrome.a comparative serologic and virologic study was performed in pigs from 5 herds with postweaning multisystemic wasting syndrome (pmws) and 2 herds without pmws in quebec. in each herd, 60 blood samples were collected at 4-wk intervals from pigs from 3 to 23 wk of age. the serum was evaluated for the presence of antibodies to porcine circovirus type 2 (pcv2) and porcine reproductive and respiratory syndrome virus (prrsv), as well as for the presence of nucleic acid of pcv2, prrsv, an porcine parvo ...200312760476
multiplex nested pcr compared with in situ hybridization for the differentiation of porcine circoviruses and porcine parvovirus from pigs with postweaning multisystemic wasting syndrome.multiplex nested polymerase chain reactions (pcrs) were developed for the simultaneous detection and differentiation of genomic material of porcine circovirus 1 (pcv1), porcine circovirus 2 (pcv2), and porcine parvovirus (ppv) in formalin-fixed, paraffin-embedded tissues. multiplex conventional and nested pcr and in situ hybridization were compared for their ability to detect the 3 viruses in such tissues. xylene deparaffinization followed by proteinase k digestion yielded dna of sufficient qual ...200312760479
ginseng extract in aluminium hydroxide adjuvanted vaccines improves the antibody response of pigs to porcine parvovirus and erysipelothrix rhusiopathiae.ginseng, the dry extract prepared from the panax ginseng c.a. mayer-root contain immunomodulators named ginsenosides, which in the pig enhance the antibody response to viral and bacterial antigens. the enhancing effect of ginseng was demonstrated vaccinating pigs against porcine parvovirus (ppv) and erysipelothrix rhusiopathiae infections, using commercially available vaccines. the potency of the licensed, aluminium hydroxide adjuvanted; vaccines were compared with those supplemented with ginsen ...200312507846
factors affecting the transfer of porcine parvovirus antibodies from sow to piglets.the aim of the study was to investigate the effects of a number of environmental, behavioural and biological factors on passive immunization of piglets as assessed by transfer of porcine parvovirus (ppv) antibodies (ab) from the colostrum of ppv vaccinated mothers to the serum of the piglets. twenty primiparous sows were housed in pens with peat, straw and branches for nest building. half the sows were prevented from achieving feedback from a completed farrowing nest by repeated removal of the n ...200212489874
the major envelope protein, gp5, of a european porcine reproductive and respiratory syndrome virus contains a neutralization epitope in its n-terminal ectodomain.a set of neutralizing monoclonal antibodies (mabs) directed against the gp(5) protein of european type porcine reproductive and respiratory syndrome virus (prrsv) has been produced previously (weiland et al., 1999). this set reacted with a plaque-purified virus (ppv) subpopulation of dutch isolate intervet-10 (i-10), but not with the european prototype prrsv lv. in order to map the neutralization epitope in the gp(5) protein of the ppv strain, the orf5 nucleotide sequence of ppv was determined. ...200312771423
serum aminotransferase levels and platelet count as predictive factor of fibrosis and cirrhosis in patients with chronic hepatitis c assess the ratio of aminotransferases in combination with platelets count as a predictive factor for stages of fibrosis in patients with chronic hepatitis c virus infection.200312779023
genetic variability of porcine parvovirus isolates revealed by analysis of partial sequences of the structural coding gene vp2.the 3'-terminal 853 nt (and the putative 283 aa) sequence of the vp2-encoding gene from 29 field strains of porcine parvovirus (ppv) were determined and compared both to each other and with other published sequences. sequences were examined using maximum-parsimony and statistical analyses for nucleotide diversity and sequence variability. among the nucleotide sequences of the ppv field strains, 26 polymorphic sites were encountered; 22 polymorphic sites were detected in the putative amino acid s ...200312771420
genetic linkage maps of two apricot cultivars ( prunus armeniaca l.), and mapping of ppv (sharka) resistance.genetic linkage maps for two apricot cultivars have been constructed using aflp, rapd, rflp and ssr markers in 81 f1 individuals from the cross 'goldrich' x 'valenciano'. this family segregated for resistance to 'plum pox virus' (ppv), the most-important virus affecting prunus species. of the 160 rapd arbitrary primers screened a total of 44 were selected. sixty one polymorphic rapd markers were scored on the mapping population: 30 heterozygous in 'goldrich', 19 heterozygous in 'valenciano', seg ...200212582518
clinical relevance of total hcv core antigen testing for hepatitis c monitoring and for predicting patients' response to study the correlation between total hepatitis c virus (hcv) core antigen (ag) and hcv-rna, and to assess the proficiency of hcv core ag testing in monitoring and predicting virologic response during and after pegylated interferon (peg-ifn) and ribavirin combination therapy. a total of 307 samples from treated and untreated patients were used to assess the correlation between the total hcv core ag test and quantitative hcv-rna assays (superquant, and quantiplex branched dna 2.0 assay). twenty- ...200312823600
expression of self-complementary hairpin rna under the control of the rolc promoter confers systemic disease resistance to plum pox virus without preventing local infection.homology-dependent selective degradation of rna, or post-transcriptional gene silencing (ptgs), is involved in several biological phenomena, including adaptative defense mechanisms against plant viruses. small interfering rnas mediate the selective degradation of target rna by guiding a multicomponent rnase. expression of self-complementary hairpin rnas within two complementary regions separated by an intron elicits ptgs with high efficiency. plum pox virus (ppv) is the etiological agent of shar ...200312823862
pathogenesis of postweaning multisystemic wasting syndrome reproduced by co-infection with korean isolates of porcine circovirus 2 and porcine parvovirus.thirty-two colostrum-deprived conventional pigs aged 28 days were co-infected with korean isolates of porcine circovirus 2 (pcv2) and porcine parvovirus (ppv) and observed for up to 35 days post-inoculation (dpi). histopathologically, granulomatous inflammation, with or without intracytoplasmic inclusion bodies, was present in lymphoid tissues (e.g., lymph nodes, thymus, spleen and tonsil) from 20 dpi, being most severe at 24 dpi. pcv2 and ppv dna were detected in the lymph nodes from 3 to 35 dp ...200312531687
ginseng and aluminium hydroxide act synergistically as vaccine adjuvants.the dry extract prepared from the panax ginseng c.a. meyer-root (total ginseng (t-ginseng)) contain ginsenosides (g-des) which were shown to have adjuvant properties as demonstrated by: (a) injecting guinea pigs with a mixture of t-ginseng and inactivated porcine parvovirus (ppv) as a conventional vaccine; (b)injecting ppv-antigen and t-ginseng simultaneously but separately at different sites on the animal and (c)injecting only the t-ginseng 1 or 2 weeks prior to immunisation with the ppv-antige ...200312559792
simultaneous detection and differentiation between porcine circovirus and porcine parvovirus in boar semen by multiplex seminested polymerase chain reaction.a multiplex seminested polymerase chain reaction (pcr) was developed for the simultaneous detection and differentiation among porcine circovirus 1 (pcv1), pcv2, and porcine parvovirus (ppv) from boar semen. primers of pcv1, pcv2 and ppv were specific and did not react with other viruses respectively. twenty (20.4%) and 42 (42.9%) out of 98 whole semen samples were found to be positive for pcv and ppv using multiplex conventional and seminested pcr, respectively. when the separated fractions of p ...200312867738
elimination of ppv and pnrsv through thermotherapy and meristem-tip culture in nectarine.the plum pox virus (ppv) and prunus necrotic ringspot virus (pnrsv) cause serious disease problems in stone-fruit trees. in this work, the possibility of obtaining plant material free from these viruses through thermotherapy and meristem-tip culture from infected nectarine shoots (prunus persica var. nectarina max, cv. 'arm king') was studied. in addition, the detection of these viruses in in vitro cultures and young acclimatized plantlets with double antibody sandwich-enzyme-linked immunosorben ...200312898177
an apricot (prunus armeniaca l.) f2 progeny linkage map based on ssr and aflp markers, mapping plum pox virus resistance and self-incompatibility traits.a genetic linkage map of apricot ( prunus armeniaca l.) was constructed using aflp and ssr markers. the map is based on an f(2) population (76 individuals) derived from self-pollination of an f(1) individual ('lito') originated from a cross between 'stark early orange' and 'tyrinthos'. this family, designated as 'lito' x 'lito', segregated for two important agronomical traits: plum pox virus resistance (ppv) and self-incompatibility. a total of 211 markers (180 aflps, 29 ssrs and two agronomic t ...200312845439
inclusion of hpv testing in routine cervical cancer screening for women above 29 years in germany: results for 8466 a prospective cohort study 8466 women attending routine cervical cancer screening were recruited. colposcopy was performed on women with any degree of atypia on cytology and/or a positive high-risk human papillomavirus (hpv)-dna test (hc2; hybrid capture 2((c))), and for a randomly selected sample of 3.4% women with negative findings on both. quality control included reviews of cytology, histology, colposcopy images and retesting of samples with polymerase chain reaction. test diagnostic perf ...200312771924
design and function of a conformationally restricted analog of the influenza virus fusion peptide.a conformationally restricted analog of the n-terminal 12-residue peptide segment of the ha2 subunit of the ppv/34, pr/8/34, and jap/57 strains of influenza virus hemagglutinin was synthesized containing three residues of calpha-methyl-valine. this peptide has a higher content of helical structure in the presence of 50% trifluoroethanol or when interacting with liposomes of egg phosphatidylcholine compared with the conformationally more flexible control peptide with the native sequence. the cont ...200312787447
effect of temperature on plum pox virus of the key factors of progress of an epidemic is the duration of virus availability for a vector in plants, which could be influenced by temperature. using five epidemiologically different isolates of plum pox virus (ppv) we studied the effect of temperature on the virus infectivity, intensity of disease symptoms and virus accumulation in nicotiana benthamiana plants as determined by a double-antibody sandwich-enzyme-linked immunosorbent assay (das-elisa). no differences in infectivity and i ...200312828345
establishment of a porcine parvovirus (ppv) dna standard and evaluation of a new lightcycler nested-pcr assay for detection of ppv.porcine parvovirus (ppv) is a major causative agent in a syndrome of reproductive failure in swine. in validation (viral clearance) studies, ppv is a model of non-enveloped viruses and is widely used instead of human parvovirus b19, which causes a variety of illnesses including erythema infectiosum (fifth disease) in children and hydrops fetalis in pregnant women. to improve the sensitivity of current pcr-based assays for detection of ppv and to standardize the quantification of ppv, we have dev ...200312821192
is hsv serology useful for the management of first episode genital herpes?first episode genital herpes simplex virus (hsv) infections can be classified into three groups, primary genital herpes (no previous exposure to hsv), non-primary first episode (igg antibody to hsv of the non-presenting type), and first episode with pre-existing igg hsv antibodies. the use of igm to classify first episode genital herpes has not been evaluated.200312902573
a comparison of the lymphocyte subpopulations of pigs experimentally infected with porcine circovirus 2 and/or parvovirus. 200312672380
exogenous porcine viruses.porcine organs, cells and tissues provide a viable source of transplants in humans, though there is some concern of public health risk from adaptation of swine infectious agents in humans. limited information is available on the public health risk of many exogenous swine viruses, and reliable and rapid diagnostic tests are available for only a few of these. the ability of several porcine viruses to cause transplacental fetal infection (parvoviruses, circoviruses, and arteriviruses), emergence or ...200312934944
virus-like particle production at low multiplicities of infection with the baculovirus insect cell system.the baculovirus insect cell expression system (bevs) was used for the production of self-forming porcine parvovirus-like particles (vlps) in serum-free medium. a low multiplicity of infection (moi) strategy was used to overcome an extra virus amplification step, undesirable in industrial production, and to minimize the virus passage effect. it was confirmed that the time of infection (toi) and moi are dependent variables. higher cell densities were obtained at low mois, keeping a constant toi; h ...200312966582
[serum antibodies detection for serological diagnosis of severe acute respiratory syndrome].to evaluate the significance of serum specific antibodies against severe acute respiratory syndrome (sars) virus in the diagnosis of sars and the concordance of ifa and elisa in the antibodies measurements.200312899765
in vivo, dendritic cells can cross-present virus-like particles using an endosome-to-cytosol pathway.recombinant parvovirus-like particles (ppv-vlps) are particulate exogenous ags that induce strong ctl response in the absence of adjuvant. in the present report to decipher the mechanisms responsible for ctl activation by such exogenous ag, we analyzed ex vivo and in vitro the mechanisms of capture and processing of ppv-vlps by dendritic cells (dcs). in vivo, ppv-vlps are very efficiently captured by cd8alpha- and cd8alpha+ dcs and then localize in late endosomes of dcs. macropinocytosis and lip ...200312928368
virus concentration using sulfonated magnetic beads to improve sensitivity in nucleic acid amplification enhance the sensitivity of virus detection by polymerase chain reaction (pcr) and reverse-transcriptional (rt)-pcr, we developed a novel virus-concentration method using sulfonated (so-) magnetic beads in the presence of divalent cations. in the presence of either zn(2+) or cu(2+) ions, we showed that so-magnetic beads were able to concentrate non-enveloped model viruses, such as porcine parvovirus (ppv) and poliovirus, which were not concentrated by polyethyleneimine (pei)-magnetic beads.(1) ...200312913251
myocarditis and abortion associated with intrauterine infection of sows with porcine circovirus 2.porcine circovirus 2 (pcv2) is a recently identified agent that has been associated with postweaning multisystemic wasting syndrome (pmws) in swine populations. in this report, the potential spectrum of disease associated with pcv2 is expanded by evidence of vertical transmission and associated reproductive failure. pcv2 was isolated from a litter of aborted piglets from a farm experiencing late-term abortions and stillbirths. severe, diffuse myocarditis was present in 1 piglet associated with e ...199912968736
association of porcine circovirus 2 with porcine respiratory disease complex.a retrospective study was performed on natural cases of porcine respiratory disease complex (prdc) to determine the association and prevalence of prdc with porcine circovirus 2 (pcv2) and other co-existing pathogens in korea. histologically, alveolar septa were markedly thickened by infiltrates of mononuclear cells. moderate to marked multifocal peribronchial and peribronchiolar fibrosis were present and often extended into the airway lamina propria. among the 105 pigs with prdc, 85 were positiv ...200314550736
detection of porcine teschoviruses and enteroviruses by lightcycler real-time pcr.porcine picornaviruses comprising at least 23 serotypes grouped into six species were described as causative agents of neurological disorders, reproductive failure, and aphthae-like dermal lesions of swine. other viruses such as classical swine fever virus (csfv), african swine fever virus, pseudorabies virus (prv), vesicular stomatitis virus, vesicular exanthema virus, porcine respiratory and reproductive syndrome virus, and porcine parvovirus (ppv) may cause diseases with similar clinical symp ...200314500127
virus concentration using polyethyleneimine-conjugated magnetic beads for improving the sensitivity of nucleic acid amplification enhance the sensitivity of virus detection by polymerase chain reaction (pcr) and reverse-transcriptional (rt)-pcr, we developed a novel virus concentration method using polyethyleneimine (pei)-conjugated magnetic beads. pei-magnetic beads adsorbed efficiently the enveloped viruses sindbis virus and herpes simplex 1 virus, and the nonenveloped virus sv-40, but not the nonenveloped viruses porcine parvovirus (ppv) or poliovirus, based on the pcr detection data. furthermore, the infectivity in ...200314599674
risk factors for porcine post-weaning multisystemic wasting syndrome (pmws) in 149 french farrow-to-finish herds.a cross-sectional study involving 149 farms was carried out in france in 2000 and 2001 to assess the risk factors for post-weaning multisystemic wasting syndrome (pmws). the farms were divided into three groups according to their current or past pmws status: cases (current and typical pmws), controls#1 (pmws-free farms), and controls#2 (farms which have recovered from pmws). two different comparisons were tested: cases versus controls#1 and cases versus controls#2. in the first comparison, the o ...200314554144
do std clinics correctly diagnose stds? an assessment of std management in hefei, china.the main purposes of the study were to assess the accuracy of laboratory testing and the diagnosis by physicians in sexually transmitted disease (std) clinics in hefei, china. among 347 men complaining of urethral discharge or dysuria, 240 tested positive at the national centre for either neisseria gonorrhoeae or chlamydia trachomatis, 310 tested positive according to the clinic laboratories, and 347 were diagnosed by the physicians. for chlamydia, the sensitivity and positive predictive value ( ...200314596769
parvovirus host range, cell tropism and evolution.the past few years have seen major advances in our understanding of the controls of evolution, host range and cell tropism of parvoviruses. notable findings have included the identification of the transferrin receptor tfr as the cell surface receptor for canine parvovirus and feline panleukopenia virus, and also the finding that specific binding to the canine tfr led to the emergence of canine parvovirus as a new pathogen in dogs. the structures of the adeno-associated virus-2 and porcine parvov ...200312941411
genomes of the parapoxviruses orf virus and bovine papular stomatitis virus.bovine papular stomatitis virus (bpsv) and orf virus (orfv), members of the genus parapoxvirus of the poxviridae, are etiologic agents of worldwide diseases affecting cattle and small ruminants, respectively. here we report the genomic sequences and comparative analysis of bpsv strain bv-ar02 and orfv strains ov-sa00, isolated from a goat, and ov-ia82, isolated from a sheep. parapoxvirus (ppv) bv-ar02, ov-sa00, and ov-ia82 genomes range in size from 134 to 139 kbp, with an average nucleotide com ...200414671098
concurrent presence of porcine circovirus type 2 and porcine parvovirus in retrospective cases of exudative epidermitis in pigs. 200414623159
whole blood samples as an alternative to serum for detection of immunity to measles virus by elisa.the aim of this study was to evaluate enzyme linked immunosorbent assay (elisa) as a testing strategy for detection of antibodies against measles virus from microquantities of blood soaked onto filter paper. we studied 165 healthy children in the age group of 1 to 2 years, attending the outpatient department of pediatrics. two sets of samples were collected from each child. one by venipuncture and the other on whatman filter paper-3 discs of 20 mm size by finger or heel prick so that each strip ...200314711476
suppressor activity of potyviral and cucumoviral infections in potyvirus-induced transgene silencing.the process known as 'recovery' by which virus-infected plants become resistant to the infection is an interesting phenomenon where both rna silencing and virus resistance fully converge. in a previous study, we showed that transgenic nicotiana benthamiana nibv3 plants, transformed with a mutated nib coding sequence from plum pox virus (ppv), showed a delayed, very specific, resistance phenotype, which was induced by the initial infection. this recovery was the consequence of the activation of a ...200313679623
fluorescent labelling reveals spatial separation of potyvirus populations in mixed infected nicotiana benthamiana plants.the distribution of potyviruses in mixed infected nicotiana benthamiana plants was investigated by using green and red fluorescent proteins (gfp, dsred). full-length cdna clones of plum pox virus (ppv-nat-agfps; ppv-nat-red), tobacco vein mottling virus (tvmv-gfp; tvmv-red) and clover yellow vein virus (clyvv-gfp) expressing fluorescent proteins, referred to here as labelled viruses, were used to characterize the distribution of different potyviral populations (e.g. tvmv-gfp/ppv-nat-red), as wel ...200313679622
interfacial enzymology of parvovirus phospholipases a2.the capsid of parvoviruses proteins were recently shown to contain secreted phospholipase a(2) (spla(2))-like activity that is required during host cell entry. parvoviral pla(2) domains have little sequence identity with spla(2)s and lack disulfide bonds. in the present study, after bacterial expression and purification, the biochemical characterizations of these first pla(2)s identified in viruses have been investigated, and a comparison has been made with other known pla(2)s. the specific acti ...200414726513
scale-up of virus-like particles production: effects of sparging, agitation and bioreactor scale on cell growth, infection kinetics and productivity.the baculovirus-insect cells expression system was used for the production of self-forming porcine parvovirus (ppv) like particles (virus-like particles, vlps) in serum-free medium. at 2l bioreactor scale an efficient production was achieved by infecting the culture at a concentration of 1.5 x 10(6)cells/ml using a low multiplicity of infection of 0.05 pfu per cell. in a continuous bioreactor, it was shown that the uninfected insect cells were not sensitive to local shear stress values up to 2.2 ...200414687971
specificity of the antibody response to the pneumococcal polysaccharide and conjugate vaccines in human immunodeficiency virus-infected adults.nonspecific antibodies, which are thought to be nonprotective, have been shown to contribute a substantial proportion of the measured concentration in the standardized immunoglobulin g (igg) enzyme-linked immunosorbent assay (elisa) for pneumococcal polysaccharide capsular antibodies. the presence of such antibodies in human immunodeficiency virus (hiv)-infected persons has not been evaluated. the amount of nonspecific antibodies is proportional to the reduction in igg antibody concentration tha ...200414715560
controlled gamma-irradiation mediated pathogen inactivation of human urokinase preparations with significant recovery of enzymatic activity.human sources of urokinase have led to the contamination of in-process lots of commercially available material with human pathogens. effective pathogen inactivation of urokinase preparations can be achieved through the use of gamma-irradiation. additionally, the presence of a free radical scavenger (ascorbate) and the control of temperature have resulted in maintenance of the enzymatic activity of urokinase without a significant effect on the pathogen inactivation properties of gamma-irradiation ...200314624796
pmws: experimental model and co-infections.porcine circovirus type 2 (pcv2) is now recognised as the causal agent of porcine multisystemic wasting syndrome (pmws), an economically important wasting disease of young pigs [j. vet. diagn. invest. 12 (2000) 3]. gross lesions of pmws include generalised lymphadenopathy, hepatitis, nephritis and pneumonia and typical histological lesions include lymphocytic depletion and multinucleated giant cell formation in lymph nodes, degeneration and necrosis of hepatocytes, and multifocal lymphohistocyti ...200414741129
the complete nucleotide sequence of plum pox virus isolates from sweet (ppv-swc) and sour (ppv-soc) cherry and their taxonomic relationships within the species.plum pox virus (ppv) sweet (swc) and sour (soc) cherry isolates were the first ppv isolates to be recovered from natural infection in sweet and sour cherry plants, respectively. their complete nucleotide sequences have been determined finding a deduced genome organisation typical for ppv species. both genomes are 9795 nucleotides long, excluding the 3' terminal poly(a) tail, and contain an open reading frame of 9432 nt, encoding a polyprotein of 3143 amino acids. the nucleotide and predicted ami ...200314579174
peracetic acid-ethanol treatment of allogeneic avital bone tissue transplants--a reliable sterilization method.based on the european standard en 1040, the validation guidelines of the german federal institute for drugs and medical devices and cpmp guidelines we tested the antimicrobial effectiveness of a peracetic acid-ethanol sterilization procedure (pes) in allogenic avital bone transplants.200314626574
porcine circovirus-2 and concurrent infections in the field.porcine circovirus-2 (pcv-2) is the necessary cause of post-weaning multisystemic wasting syndrome (pmws) in swine; however, a variety of co-factors, including other infectious agents, are thought to be necessary in the full expression of disease. porcine parvovirus (ppv) was found in the inoculum used in the first experiments to reproduce pmws in gnotobiotic swine. retrospective and prospective studies in the field and laboratory have demonstrated pcv-2 can act synergistically with ppv to enhan ...200414741128
recombinant protein-based enzyme-linked immunosorbent assay and immunochromatographic tests for detection of immunoglobulin g antibodies to severe acute respiratory syndrome (sars) coronavirus in sars enzyme-linked immunosorbent assay (elisa) and a rapid immunochromatographic test for detection of immunoglobulin g (igg) antibodies in severe acute respiratory syndrome (sars) patients were developed by utilizing the well-characterized recombinant proteins gst-n and gst-u274. the elisa detected igg antibodies to sars-cov in all 74 convalescent-phase samples from sars patients while weakly cross-reacting to only 1 of the 210 control sera from healthy donors. this finding thus led to a kit sens ...200415013977
photochemical inactivation of selected viruses and bacteria in platelet concentrates using riboflavin and light.a medical device is being developed for the reduction of pathogens in plt concentrates (pcs). the device uses broadband uv light and the compound riboflavin (vitamin b(2)).200415157255
a comparison of virus isolation, polymerase chain reaction, immunohistochemistry, and in situ hybridization for the detection of porcine circovirus 2 and porcine parvovirus in experimentally and naturally coinfected pigs.virus isolation, polymerase chain reaction (pcr), immunohistochemistry, and in situ hybridization were compared for the detection of porcine circovirus 2 (pcv2) and porcine parvovirus (ppv) from experimentally and naturally coinfected pigs. all coinfected pigs developed postweaning multisystemic wasting syndrome (pmws), characterized by sudden onset of depression and anorexia. microscopically, granulomatous inflammation with intracytoplasmic inclusion bodies was present in lymph node from all co ...200414974846
influence of flanking sequences on presentation efficiency of a cd8+ cytotoxic t-cell epitope delivered by parvovirus-like particles.we have previously developed an antigen-delivery system based on hybrid recombinant porcine parvovirus-like particles (ppv-vlps) formed by the self-assembly of the vp2 protein of ppv carrying a foreign epitope at its n terminus. in this study, different constructs were made containing a cd8(+) t-cell epitope of chicken ovalbumin (ova) to analyse the influence of the sequence inserted into vp2 on the correct processing of vlps by antigen-presenting cells. we analysed the presentation of the ova e ...200414993639
development of an elisa based on the baculovirus-expressed capsid protein of porcine circovirus type 2 as antigen.the genome of porcine circovirus type 2 (pcv2) contains two major open reading frames, which have been shown to encode the virus capsid and replication-associated proteins. the capsid protein is a major structural protein of the virus; it can be a suitable target antigen for detecting pcv2-specific antibodies to monitor pcv2 infection. to produce the antigen, the capsid protein coding sequence was cloned into a baculovirus transfer vector, and a recombinant capsid (rc) protein of pcv2 was expres ...200415107550
hiv and hepatitis c coinfection within the caesar study.the declining incidence of aids-related opportunistic diseases among people with hiv infection has shifted the focus of clinical management to prevention and treatment of comorbidities such as chronic liver disease. the increased risk of hepatitis c virus (hcv)-related advanced liver disease in people with hiv infection makes early hcv diagnosis a priority. to assess hcv prevalence and predictors of hiv/hcv coinfection, we have conducted a retrospective analysis of people enrolled in the caesar ...200415139984
multiplex pcr for rapid detection of pseudorabies virus, porcine parvovirus and porcine circoviruses.a multiplex pcr (mpcr) assay was developed and subsequently evaluated for its effectiveness as a means to simultaneously detect multiple viral infections of swine. specific primers for each of four common dna viruses, namely, pseudorabies virus (prv), porcine circovirus type i (pcv1), porcine circovirus type ii (pcv2), and porcine parvovirus (ppv), were used for testing procedure. the assay was shown to be highly sensitive in that as little as 10(-4) ng of each of the respective amplicons (appro ...200415223125
removal of small non-enveloped viruses by nanofiltration.nanofiltration is one of the most effective virus reduction methods in the manufacturing process of plasma products. however, it is difficult to remove small viruses from high molecular weight protein preparations like immunoglobulin g or factor viii complex by nanofiltration, because the size of the protein is similar to that of viruses. in order to separate the viruses from these proteins by nanofiltration, it is necessary to change the size of either one. in this study, we report that such no ...200415144526
specific detection and quantification of plum pox virus by real-time fluorescent reverse transcription-pcr.plum pox virus (ppv), a destructive and economically devastating pathogen of prunus species, was recently discovered in pennsylvania and canada. current containment efforts involve eradication of infected trees based on elisa surveys, which are laborious and less sensitive than pcr-based techniques. a real-time, fluorescent, reverse transcription-polymerase chain reaction (rt-pcr) assay was developed for the detection of ppv in the smart cycler (cepheid). the methods developed are reproducible, ...200415234814
a polysaccharide fraction from medicinal herb prunella vulgaris downregulates the expression of herpes simplex virus antigen in vero cells.herpes simplex viruses (hsv) are pathogenic. with the emergence of drug-resistant strains of hsv, new antiviral agents, especially those with different modes of action, are urgently needed. prunella vulgaris l. (labiatae), a perennial plant commonly found in china and europe, has long been used as a folk medicine to cure ailments. in this study, a polysaccharide fraction was prepared from prunella vulgaris (ppv), and its effects on the expressions of hsv-1 and hsv-2 antigens in their host vero c ...200415182906
analysis and comparison of the 4b core protein gene of avipoxviruses from wild birds: evidence for interspecies spatial phylogenetic variation.avipoxviruses have been isolated from a wide variety of avian hosts, and yet little is known regarding the host-virus species variation of the genus avipoxvirus. we have investigated the variations in the viral 4b core protein gene from six different avipoxviruses based on pcr, southern blot and nucleotide sequence analysis to evaluate the suitability of this region for differentiation between avipoxvirus isolates. southern blot and nucleotide sequence analysis revealed considerable interspecies ...200415290371
effect of porcine parvovirus vaccination on the development of pmws in segregated early weaned pigs coinfected with type 2 porcine circovirus and porcine parvovirus.the objectives of this study were to determine if coinfection of segregated early weaned (sew) pigs with porcine circovirus type 2 (pcv2) and porcine parvovirus (ppv) induces an increase in the incidence of post-weaning multisystemic wasting syndrome (pmws) compared to singular pcv2 infection, and to determine if vaccination against ppv protects pigs against pmws associated with pcv2/ppv coinfection in sew pigs. seventy, 3-week-old, sew pigs were randomly assigned to one of the five groups. pigs ...200415036529
quantification of epstein-barr virus load in argentinean transplant recipients using real-time lymphoproliferative disease (ptld) is a frequent and severe epstein-barr virus (ebv)-associated complication in transplant recipients that is caused by suppression of t-cell function.200415288615
comparison of the efficacy of virus inactivation methods in allogeneic avital bone tissue transplants.several procedures for inactivating viruses are used presently in the context of bone tissue transplants. common methods used are gamma irradiation (25 kgy), treatment with moist heat (82.5 degrees c/15 min., lobator-sd2-system) as well as chemical sterilisation using peracetic acid-ethanol treatment (pes, 2% peracetic acid, 96% ethanol, aqua [2:1:1], 200 mbar, agitation, 4 hours). based on national and international guidelines, we tested the antivirucidal effectiveness of these methods in human ...200115256903
geographically and temporally distant natural recombinant isolates of plum pox virus (ppv) are genetically very similar and form a unique ppv subgroup.natural recombinant plum pox virus (ppv) isolates were detected in albania, bulgaria, czech republic, germany, hungary and slovakia. despite different geographical origins and dates of isolation, all the recombinant isolates were closely related at the molecular level and shared the same recombination breakpoint as well as a typical signature in their n-terminal coat protein sequence, suggesting a common origin. biological assays with four recombinant isolates demonstrated their capacity to be a ...200415302961
experimental squalene adjuvant. i. preparation and testing of its effectiveness.the authors prepared experimental lipoid adjuvant of the oil-in-water (o/w) type, based on squalene (esa: experimental squalene adjuvant), intended for liquid inactivated veterinary vaccines. the final concentration of squalene in the vaccine was 5% (w/v). detergents poloxamer 105 in the concentration of 4% (w/v) and abil-care in the concentration of 2% were used as emulsifiers. the water component was inactivated virus suspension. emulsification at decreased temperatures (up to 37 degrees c) fa ...200415308373
human papillomavirus (hpv) testing in the management of women with abnormal pap smears. experience of a colposcopy referral clinic.several detailed algorithms for the appropriate use of human papillomavirus (hpv) testing in the management of women with abnormal pap (papanicolaou) smears have been launched, but their direct country-to-country adoption is difficult. this necessitates their testing in individual settings, which is ongoing in our colposcopy referral clinic.200415493169
serum levels of hepatitis c virus core antigen as a marker of infection and response to therapy.hepatitis c virus (hcv) core antigen is a recently developed marker of hepatitis c infection. we compared the predictive power of hcv core antigen with reverse transcription polymerase chain reaction (rt-pcr) and branched dna assay for hcv-rna as markers of infection and response to interferon therapy.200415330912
inhibitory effects of cystatins on proteolytic activities of the plum pox potyvirus cysteine an effort to develop new antiviral strategies effective against potyviruses, several cystatins were evaluated for their ability to inhibit the cysteine proteinases of plum pox potyvirus (ppv) using in vitro proteolytic assays. the following cystatins were purified as gst fusion proteins and shown to be active against papain:oryzacystatins i and ii (oci and ocii), corn cystatin ii (ccii), human stefin a (hsa), the domain 8 of tomato multicystatin (tmc-8) and a large 24kda tomato cystatin (ltcy ...200415351491
p16(ink4a) expression is related to grade of cin and high-risk human papillomavirus but does not predict virus clearance after conization or disease outcome.the role of p16(ink4a) as a marker of hr-hpv and in the diagnosis of cin has been well established, but its predictive value in the clearance of the virus after cin treatment and its use as a prognostic marker of cervical cancer has not been studied. a series of 302 archival samples, including 150 squamous cell carcinomas (sccs) and 152 cin lesions, were subjected to immunohistochemical staining for p16(ink4a) and hpv testing using pcr with three primer sets (my09/11, gp5/gp6, spf). follow-up da ...200415381905
the predictive value of p16(ink4a) and hybrid capture 2 human papillomavirus testing for high-grade cervical intraepithelial neoplasia.we performed p16(ink4a) immunocytochemical analysis and hybrid capture 2 (hc2; digene, gaithersburg, md) high-risk hpv testing on 210 abnormal surepath (tripath imaging, burlington, nc) papanicolaou specimens diagnosed as low-grade squamous intraepithelial lesion (lsil) or high grade squamous intraepithelial lesion (hsil). the results were compared with 121 follow-up biopsy specimens. p16(ink4a) was positive in 57.9% of women with lsil compared with 97.1% of women with hsil. in contrast, hc2 tes ...200415539381
stability of gene silencing-based resistance to plum pox virus in transgenic plum (prunus domestica l.) under field conditions.plum pox virus (ppv) is one of the most devastating diseases of prunus species. since few sources of resistance to ppv have been identified, transgene-based resistance offers a complementary approach to developing ppv-resistant stone fruit cultivars. c5, a transgenic clone of prunus domestica l., containing the ppv coat protein (cp) gene, has been described as highly resistant to ppv in greenhouse tests, displaying characteristics typical of post-transcriptional gene silencing (ptgs). we show in ...200415587267
a simplified rt-pcr-based detection of recombinant plum pox virus isolates.closely related natural plum pox virus (ppv) isolates derived from homologous rna recombination between ppv-d and ppv-m have been recently identified and shown naturally spread in several european countries. as their serological properties were identical with those of conventional ppv-m isolates, they could be detected only by combined analysis of at least two different genome parts. to simplify the detection of such recombinants primers specific to ppv-m and ppv-d sequences with binding sites l ...200415595211
high porcine parvovirus antibodies in sow herds: prevalence and associated factors.porcine parvovirus (ppv) is widespread among swine. our objective was to determine the prevalence of loosely housed sow herds in finland with at least one animal with high (infection level) ppv antibodies and to gather basic knowledge about vaccination practices. in addition, selected factors associated with high antibody levels found in sows were examined. altogether, 247 animals were sampled in 21 randomly chosen loosely housed sow herds. samples were analysed with the haemagglutination inhibi ...200515655002
simultaneous detection of six stone fruit viruses by non-isotopic molecular hybridization using a unique riboprobe or 'polyprobe'.a new strategy for the simultaneous detection of plant viruses by molecular hybridization has been developed. two, four or six viral sequences were fused in tandem and transcribed to render unique riboprobes and designated as 'polyprobes'. the 'polyprobe four' (poly 4) covered the four ilarviruses affecting stone fruit trees including apple mosaic virus (apmv), prunus necrotic ringspot virus (pnrsv), prune dwarf virus (pdv), and american plum line pattern virus (aplpv) whereas the 'polyprobe two ...200415664050
detection and differentiation of plum pox virus using real-time multiplex pcr with sybr green and melting curve analysis: a rapid method for strain typing.a real-time multiplex pcr procedure with melting curve analysis, using the green fluorescence dye sybr green i, was developed for rapid and reliable identification of plum pox virus (ppv) isolates of strains d and m. members of the different strains were identified by their distinctive melting temperatures (t(m)s); 84.3-84.43 degrees c for d isolates, and 85.34-86.11 degrees c for m isolates. the associated amplicon sizes were 114 and 380 bp, respectively. the procedure was used for detection an ...200515620404
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