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an extremely diverse cd4 response to vaccinia virus in humans is revealed by proteome-wide t-cell profiling.cd4 t cells are required for the maintenance and recall of antiviral cd8 t cells and for antibody responses. little is known concerning the overall architecture of the cd4 response to complex microbial pathogens. in a whole-proteome approach, 180 predicted open reading frames (orfs) in the vaccinia virus genome were expressed and tested using responder cells from 20 blood samples from 11 vaccinees. validation assays established the sensitivity and specificity of the system. overall, cd4 response ...200818480455
heterogeneity in the a33 protein impacts the cross-protective efficacy of a candidate smallpox dna vaccine.we previously developed a gene-based vaccine, termed 4pox, which targets four orthopoxvirus proteins (a33, l1, b5, and a27). because any subunit orthopoxvirus vaccine must protect against multiple species of orthopoxviruses, we are interested in understanding the cross-protective potential of our 4pox vaccine target immunogens. in our current studies, we focused on the a33 immunogen. we found one monoclonal antibody against a33, mab-1g10, which could not bind the monkeypox virus a33 ortholog, a3 ...200818482742
targeting the vaccinia virus l1 protein to the cell surface enhances production of neutralizing antibodies.the current live-orthopoxvirus vaccine is associated with minor to serious adverse affects, and is contraindicated for use in a significant portion of the population. as an alternative vaccine, we have previously shown that a dna subunit vaccine (4pox) based on four orthopoxvirus immunogens (l1r, b5r, a27l and a33r) can produce protective immunity against lethal orthopoxvirus challenges in mice and nonhuman primates. because antibodies are critical for protection against secondary orthopoxvirus ...200818485547
specific inhibition of orthopoxvirus replication by a small interfering rna targeting the d5r gene.concerns about the potential use of smallpox in bioterrorism have stimulated interest in the development of novel antiviral treatments. currently, there are no effective therapies against smallpox and new treatment strategies are greatly needed.200818572748
poxvirus host range genes.as a family of viruses, poxviruses collectively exhibit a broad host range and most of the individual members are capable of replicating in a wide array of cell types from various host species, at least in vitro. at the cellular level, poxvirus tropism is dependent not upon specific cell surface receptors, but rather upon: (1) the ability of the cell to provide intracellular complementing factors needed for productive virus replication, and (2) the ability of the specific virus to successfully m ...200818585528
real-time pcr assay to identify variants of vaccinia virus: implications for the diagnosis of bovine vaccinia in brazil.naturally occurring infections of vaccinia virus (vacv) have been recognized in brazil during the past 10 years. human brazilian vaccinia virus (bvv) infections typically occur as a zoonosis transferred from affected dairy cows to their handlers. outbreaks have caused notable economic losses to the rural community in the region. the origins of bvv are unclear but previous analyses have shown that at least two distinct clades of bvv exist. the aim of this study was to develop a rapid and inexpens ...200818602170
monkeypox virus viral chemokine inhibitor (mpv vcci), a potent inhibitor of rhesus macrophage inflammatory protein-1.monkeypox virus (mpv) is an orthopoxvirus with considerable homology to variola major, the etiologic agent of smallpox. although smallpox was eradicated in 1976, the outbreak of mpv in the u.s. highlights the health hazards associated with zoonotic infections. like other orthopoxviruses, mpv encodes a secreted chemokine binding protein, vcci that is abundantly expressed and secreted from mpv infected cells. emsa data shows vcci efficiently binds rhesus mip-1alpha (rhmip-1alpha) at near one to on ...200818639466
dual-probe real-time pcr assay for detection of variola or other orthopoxviruses with dried reagents.a real-time, multiplexed polymerase chain reaction (pcr) assay based on dried pcr reagents was developed. only variola virus could be specifically detected by a fam (6-carboxyfluorescein)-labeled probe while camelpox, cowpox, monkeypox and vaccinia viruses could be detected by a tet (6-carboxytetramethylrhodamine)-labeled probe in a single pcr reaction. approximately 25 copies of cloned variola virus dna and 50 copies of genomic orthopoxviruses dna could be detected with high reproducibility. th ...200818725245
[smallpox is a dormant volcano].the presence of rodent-associated natural foci containing at least 6 of the known 11 viruses belonging to the genus orthopoxvirus (poxviridae, chordopoxvirinae) within the equatorial, tropical, subtropical, temperate, and subarctic climatic zones; the increasing aggravation of the monkey pox epidemic situation in equatorial africa with an increase in human mortality by an average of 9.8% with a possibility of 2 to 8 passages in 30-70% of patients; the possible persistence of a virus in the human ...200818756808
the vaccinia virus fusion inhibitor proteins spi-3 (k2) and ha (a56) expressed by infected cells reduce the entry of superinfecting virus.the orthopoxvirus spi-3 (k2) and a56 (hemagglutinin, ha) proteins interact and together prevent cell-cell fusion. spi-3/a56 has been proposed to prevent the superinfection of previously infected cells by reducing virus-cell fusion. binding of mature virions of vaccinia virus (vv) to vv-infected cells was unaffected by spi-3 or a56 on the surface of infected cells. entry of vv into infected cells was assessed using vv-p(t7)-luc carrying the luciferase reporter under t7 control. cells infected wit ...200818760436
mhc-i-restricted epitopes conserved among variola and other related orthopoxviruses are recognized by t cells 30 years after vaccination.it is many years since the general population has been vaccinated against smallpox virus. here, we report that human leukocyte antigen (hla) class i restricted t cell epitopes can be recognized more than 30 years after vaccination. using bioinformatic methods, we predicted 177 potential cytotoxic t lymphocyte epitopes. eight epitopes were confirmed to stimulate ifn-gamma release by t cells in smallpox-vaccinated subjects. the epitopes were restricted by five supertypes (hla-a1, -a2, -a24 -a26 an ...200818797815
[molecular evolution of poxviruses].previous restriction fragment length polymorphism analysis divided variola virus (varv) strains into two subtypes, one of which included west african and south american isolates. this allowed a dating to be introduced for the first time in estimation of the varv evolution rate. the results were used to analyze the molecular evolution of the total family poxviridae. comparisons of the known nucleotide sequences were performed for the extended conserved central genome region in 42 orthopoxvirus st ...200818825952
potential effect of prior raccoonpox virus infection in raccoons on vaccinia-based rabies immunization.the usda, wildlife services cooperative oral rabies vaccination (orv) program uses a live vaccinia virus-vectored (genus orthopoxvirus) vaccine, raboral v-rg (v-rg), to vaccinate specific wildlife species against rabies virus in several regions of the u.s. several naturally occurring orthopoxviruses have been found in north america, including one isolated from asymptomatic raccoons (procyon lotor). the effect of naturally occurring antibodies to orthopoxviruses on successful v-rg vaccination in ...200818834520
monkeypox virus and insights into its immunomodulatory proteins.summary: monkeypox is a disease that is endemic in central and western africa. however, in 2003, there was an outbreak in the united states, representing the first documented monkeypox cases in the western hemisphere. although monkeypox virus is less fatal and not as transmissible as variola virus, the causative agent of smallpox, there is concern that monkeypox virus could become a more efficient human pathogen. the reason for this may lie in the virus' genetic makeup, ecological changes, chang ...200818837778
vaccinia virus a26 and a27 proteins form a stable complex tethered to mature virions by association with the a17 transmembrane protein.during vaccinia virus replication, mature virions (mvs) are wrapped with cellular membranes, transported to the periphery, and exported as extracellular virions (evs) that mediate spread. the a26 protein is unusual in that it is present in mvs but not evs. this distribution led to a proposal that a26 negatively regulates wrapping. a26 also has roles in the attachment of mvs to the cell surface and incorporation of mvs into proteinaceous a-type inclusions in some orthopoxvirus species. however, a ...200818842719
smallpox vaccines for biodefense: need and feasibility.smallpox, eradicated as a cause of natural disease through an intensive global effort in the later part of the 20th century, has resurfaced as a possible agent of bioterrorism. for this reason, there is renewed interest in smallpox vaccines. live vaccinia virus, an orthopoxvirus related to smallpox, has a long and successful clinical track record as an effective smallpox vaccine; however, its use is associated with uncommon yet serious adverse events. this has led to a surge of recent research i ...200818844596
poxvirus k7 protein adopts a bcl-2 fold: biochemical mapping of its interactions with human dead box rna helicase ddx3.poxviruses have evolved numerous strategies to evade host innate immunity. vaccinia virus k7 is a 149-residue protein with previously unknown structure that is highly conserved in the orthopoxvirus family. k7 bears sequence and functional similarities to a52, which interacts with interleukin receptor-associated kinase 2 and tumor necrosis factor receptor-associated factor 6 to suppress nuclear factor kappab activation and to stimulate the secretion of the anti-inflammatory cytokine interleukin-1 ...200918845156
vaccinia virus: shedding and horizontal transmission in a murine model.vaccinia virus (vacv) has been associated with several bovine vaccinia outbreaks in brazil, affecting cattle and humans. there are no available data about vacv environmental circulation or the role of wildlife in the emergence of an outbreak. since vacv was isolated from rodents in brazil, we investigated shedding and transmission of vacv strains in mice. the vacv excretion profile was assessed by pcr and chicken chorioallantoic membrane infection, revealing viral dna and infectious virus in the ...200819008383
[human infection by the cowpox virus in the microregion of itajubá, state of minas gerais: case report].the authors report three human cases of cowpox infection, among farm workers who were manually milking infected cows, in the microregion of itajubá, minas gerais. the diagnostic techniques used were: isolation of samples similar to the vaccinia virus, from skin lesion secretions; polymerase chain reaction; electronic microscopy; and antibodies for orthopoxvirus in the patients' blood.200819009196
postexposure immunization with modified vaccinia virus ankara or conventional lister vaccine provides solid protection in a murine model of human smallpox.decades after the cessation of smallpox vaccination, the potential of the deliberate release of pathogenic orthopoxviruses has forced a reconsideration of using these extremely efficient human vaccines. scenarios of sudden biothreats have prompted demand for rapidly protective vaccination. however, the feasibility of short-term vaccination (i.e., vaccination shortly before exposure) with vaccinia virus (vacv) is uncertain.200919012492
progress in the discovery of compounds inhibiting orthopoxviruses in animal models.surrogate animal models must be used for testing antiviral agents against variola (smallpox) virus infections. once developed, these, compounds can be stockpiled for use in the event of a bioterrorist incident involving either variola or monkeypox virus, or used to treat an occasional serious orthopoxvirus infection, such as disseminated vaccinia complication following exposure to the live virus vaccine. recently, considerable progress has been made in the discovery of novel antiviral agents fou ...200819024628
eczema vaccinatum resulting from the transmission of vaccinia virus from a smallpox vaccinee: an investigation of potential fomites in the home environment.on march 3, 2007, a 2-year-old boy was hospitalized with eczema vaccinatum. his two siblings, one with eczema, were subsequently removed from the home. swabs of household items obtained on march 13th were analyzed for orthopoxvirus dna signatures with real-time pcr. virus culture was attempted on positive specimens. eight of 25 household samples were positive by pcr for orthopoxvirus; of these, three yielded viable vaccinia virus in culture. both siblings were found to have serologic evidence of ...200919027813
activities of certain 5-substituted 4'-thiopyrimidine nucleosides against orthopoxvirus infections.as part of a program to identify new compounds that have activity against orthopoxviruses, a number of 4'-thionucleosides were synthesized and evaluated for their efficacies against vaccinia and cowpox viruses. seven compounds that were active at about 1 microm against both viruses in human cells but that did not have significant toxicity were identified. the 5-iodo analog, 1-(2-deoxy-4-thio-beta-d-ribofuranosyl)-5-iodouracil (4'-thioidu), was selected as a representative molecule; and this comp ...200919029322
detection of human orthopoxvirus infections and differentiation of smallpox virus with real-time pcr.we developed a real-time pcr protocol to detect orthopoxviruses (opvs) from different clinical specimens and to separate variola virus from other opvs. in our protocol, we used automated nucleic acid extraction system together with real-time pcr to create a simple, safe and fast procedure to obtain an initial result. the sensitivity was better by using designed hybridization probes as compared to sybr green i for detection. the detection limit ranged from 13 to 1,300 copies per 20 microl reactio ...200919031452
inhibition of complement and cd14 attenuates the escherichia coli-induced inflammatory response in porcine whole blood.the innate immune response is a double-edged sword in systemic inflammation and sepsis. uncontrolled or inappropriate activation can damage and be lethal to the host. several studies have investigated inhibition of downstream mediators, including tumor necrosis factor alpha (tnf-alpha) and interleukin-1beta (il-1beta). emerging evidence indicates that upstream inhibition is a better therapeutic approach for attenuating damaging immune activation. therefore, we investigated inhibition of two cent ...200919047409
in vitro efficacy of st246 against smallpox and monkeypox.since the eradication of smallpox and the cessation of routine childhood vaccination for smallpox, the proportion of the world's population susceptible to infection with orthopoxviruses, such as variola virus (the causative agent of smallpox) and monkeypox virus, has grown substantially. in the united states, the only vaccines for smallpox licensed by the food and drug administration (fda) have been live virus vaccines. unfortunately, a substantial number of people cannot receive live virus vacc ...200919075062
mousepox in the c57bl/6 strain provides an improved model for evaluating anti-poxvirus therapies.the intranasal lethal mousepox model employing the a/ncr mouse strain is used to evaluate anti-orthopoxvirus therapies. these infections mimic large droplet transmission and result in 100% mortality within 7-10 days with as little as 1 pfu of ectromelia virus. unlike the a/ncr model, humans are less susceptible to lethal respiratory infections with variola virus and monkeypox virus as demonstrated by their lower mortality rates. in this study we show that a low dose intranasal infection of c57bl ...200919100593
structural basis for antagonism of human interleukin 18 by poxvirus interleukin 18-binding protein.human interleukin-18 (hil-18) is a cytokine that plays an important role in inflammation and host defense against microbes. its activity is regulated in vivo by a naturally occurring antagonist, the human il-18-binding protein (il-18bp). functional homologs of human il-18bp are encoded by all orthopoxviruses, including variola virus, the causative agent of smallpox. they contribute to virulence by suppressing il-18-mediated immune responses. here, we describe the 2.0-a resolution crystal structu ...200819104048
orthopoxviruses require a functional ubiquitin-proteasome system for productive replication.cellular homeostasis depends on an intricate balance of protein expression and degradation. the ubiquitin-proteasome pathway plays a crucial role in specifically targeting proteins tagged with ubiquitin for destruction. this degradation can be effectively blocked by both chemically synthesized and natural proteasome inhibitors. poxviruses encode a number of proteins that exploit the ubiquitin-proteasome system, including virally encoded ubiquitin molecules and ubiquitin ligases, as well as btb/k ...200919109393
a prairie dog animal model of systemic orthopoxvirus disease using west african and congo basin strains of monkeypox virus.multiple monkeypox virus (mpxv) animal models have been discussed in previous studies, but no small animal models, nor most non-human primate models, demonstrated the protracted asymptomatic incubation phase seen in systemic human orthopoxvirus illness. herein, we characterize a black-tailed prairie dog (pd) (cynomys ludovicianus) model of infection, via intranasal and intradermal exposures, with the two mpxv clades. daily observations of the animals were made (food consumption, general symptoms ...200919141441
molecule of the month. tecovirimat. 200819180269
specific targeting of the f13l protein by st-246 affects orthopoxvirus production differently.st-246 is a potent anti-orthopoxviral molecule targeting the f13l protein of vaccinia virus, which is involved in the wrapping of viruses. the discrepancy in sensitivities of several orthopoxviruses to st-246 has raised questions about potential differences in their replicative cycles and/or the presence of another drug target.200819195323
effective post-exposure protection against lethal orthopoxviruses infection by vaccinia immune globulin involves induction of adaptive immune response.the therapeutic potential of human vaccinia immunoglobulin (vig) in orthopoxvirus infection was examined using two mouse models for human poxvirus, based on ectromelia virus and vaccinia western reserve (wr) respiratory infections. despite the relatively fast clearance of human vig from mice circulation, a single vig injection protected immune-competent mice against both infections. full protection against lethal ectromelia virus infection was achieved by vig injection up to one day post-exposur ...200919195492
st-246 antiviral efficacy in a nonhuman primate monkeypox model: determination of the minimal effective dose and human dose justification.therapeutics for the treatment of pathogenic orthopoxvirus infections are being sought. in the absence of patients with disease, animal models of orthopoxvirus disease are essential for evaluation of the efficacies of antiviral drugs and establishment of the appropriate dose and duration of human therapy. infection of nonhuman primates (nhp) by the intravenous injection of monkeypox virus has been used to evaluate a promising therapeutic drug candidate, st-246. st-246 administered at 3 days post ...200919223621
cat-to-human orthopoxvirus transmission, northeastern italy. 200919239778
smallpox virus plaque phenotypes: genetic, geographical and case fatality relationships.smallpox (infection with orthopoxvirus variola) remains a feared illness more than 25 years after its eradication. historically, case-fatality rates (cfrs) varied between outbreaks (<1 to approximately 40 %), the reasons for which are incompletely understood. the extracellular enveloped virus (eev) form of orthopoxvirus progeny is hypothesized to disseminate infection. investigations with the closely related orthopoxvirus vaccinia have associated increased comet formation (eev production) with i ...200919264626
a polymerase chain reaction strategy for the diagnosis of camelpox.camelpox is a contagious viral skin disease that is mostly seen in young camels. the disease is caused by the camelpox virus (cmlv). in the present study, a polymerase chain reaction (pcr) assay based on the c18l gene (encoding ankyrin repeat protein) and a duplex pcr based on the c18l and dna polymerase (dna pol) genes were developed. the former assay yields a specific amplicon of 243 bp of the c18l gene, whereas the duplex pcr yields 243- and 96-bp products of the c18l and dna pol genes, respe ...200919286503
the octadecyloxyethyl ester of (s)-9-[3-hydroxy-2-(phosphonomethoxy) propyl]adenine is a potent and selective inhibitor of hepatitis c virus replication in genotype 1a, 1b, and 2a replicons.the octadecyloxyethyl (ode) and hexadecyloxypropyl (hdp) esters of (s)-9-[3-hydroxy-2-(phosphonomethoxy)propyl]adenine (hpmpa) are potent inhibitors of orthopoxvirus, herpesvirus, human immunodeficiency virus type 1, and hepatitis b virus replication in vitro. hdp and ode esters of (s)-hpmpa and (r)-hpmpa were evaluated for their activity in hepatitis c virus (hcv) replicon assays using luciferase (1b and 2a replicons) or rna (1b) quantification. the ode ester of (s)-hpmpa [ode-(s)-hpmpa] was th ...200919289518
inhibition of vaccinia virus replication by two small interfering rnas targeting b1r and g7l genes and their synergistic combination with cidofovir.in view of the threat of the potential use of variola virus in a terrorist attack, considerable efforts have been performed to develop new antiviral strategies against orthopoxviruses. here we report on the use of rna interference, either alone or in combination with cidofovir, as an approach to inhibit orthopoxvirus replication. two selected small interfering rnas (sirnas), named sib1r-2 and sig7l-1, and a previously reported sirna, i.e., sid5r-2 (which targets the viral d5r mrna), were evaluat ...200919307376
differentiation of variola major and variola minor variants by mgb-eclipse probe melt curves and genotyping analysis.smallpox, caused by the variola major virus, is considered to be one of the most lethal of all potential biological weapons and has far-reaching consequences. real-time polymerase chain reaction (pcr) assays are available as a reliable diagnostic tool to detect members of the genus orthopoxvirus. in addition real-time pcr assays specific for the variola virus have been developed that distinguish it from other orthopoxviruses. however, a positive identification of variola spp. does not classify t ...200919345728
monkeypox or varicella? lessons from a rash outbreak investigation in the republic of the congo.monkeypox virus and varicellazoster virus (vzv) cause visually similar rash illnesses. monkeypox is more virulent, with fatality rates up to 10%. in june 2007, reports were received of a rash illness outbreak in isolated villages in likouala district, republic of the congo. blood specimens were obtained from 142 individuals reporting rash illness between january and september 2007 from four villages in likouala. thirty-seven cases of probable vzv were identified based on low vzv igg avidity; cas ...200919346366
nonhuman primates are protected from smallpox virus or monkeypox virus challenges by the antiviral drug st-246.st-246, a potent orthopoxvirus egress inhibitor, is safe and effective at preventing disease and death in studies of small-animal models involving challenge by several different pathogenic poxviruses. in this report, the antiviral efficacy of st-246 in treatment of nonhuman primates infected with variola virus or monkeypox virus was assessed. the data indicate that oral dosing once per day with st-246 protects animals from poxvirus disease, as measured by reductions in viral load and numbers of ...200919349521
the orthopoxvirus 68-kilodalton ankyrin-like protein is essential for dna replication and complete gene expression of modified vaccinia virus ankara in nonpermissive human and murine cells.modified vaccinia virus ankara (mva) is a highly attenuated and replication-deficient vaccinia virus (vacv) that is being evaluated as replacement smallpox vaccine and candidate viral vector. mva lacks many genes associated with virulence and/or regulation of virus tropism. the 68-kda ankyrin-like protein (68k-ank) is the only ankyrin repeat-containing protein that is encoded by the mva genome and is highly conserved throughout the orthopoxvirus genus. we showed previously that 68k-ank is compos ...200919357172
activation of the pi3k/akt pathway early during vaccinia and cowpox virus infections is required for both host survival and viral replication.viral manipulation of the transduction pathways associated with key cellular functions such as actin remodeling, microtubule stabilization, and survival may favor a productive viral infection. here we show that consistent with the vaccinia virus (vacv) and cowpox virus (cpxv) requirement for cytoskeleton alterations early during the infection cycle, pbk/akt was phosphorylated at s473 [akt(s473-p)], a modification associated with the mammalian target of rapamycin complex 2 (mtorc2), which was par ...200919386722
expanding the repertoire of modified vaccinia ankara-based vaccine vectors via genetic complementation strategies.modified vaccinia virus ankara (mva) is a safe, highly attenuated orthopoxvirus that is being developed as a recombinant vaccine vector for immunization against a number of infectious diseases and cancers. however, the expression by mva vectors of large numbers of poxvirus antigens, which display immunodominance over vectored antigens-of-interest for the priming of t cell responses, and the induction of vector-neutralizing antibodies, which curtail the efficacy of subsequent booster immunization ...200919421328
inhibition of vaccinia virus replication by peptide aptamers.a20 protein is a major component of the vaccinia virus replication complex. it binds to the dna polymerase e9, the uracil dna glycosylase d4 and the primase/helicase d5, three proteins that are essential for viral dna synthesis. the identification of molecules able to interact with the replication complex and inhibit its activity is a promising strategy for the design of new anti-orthopoxvirus drugs. in this study, we used a yeast genetic approach to select, from combinatorial libraries, 8-mers ...200919428604
proteomic screening of variola virus reveals a unique nf-kappab inhibitor that is highly conserved among pathogenic orthopoxviruses.identification of the binary interactions between viral and host proteins has become a valuable tool for investigating viral tropism and pathogenesis. here, we present the first systematic protein interaction screening of the unique variola virus proteome by using yeast 2-hybrid screening against a variety of human cdna libraries. several protein-protein interactions were identified, including an interaction between variola g1r, an ankryin/f-box containing protein, and human nuclear factor kappa ...200919451633
detection of camel pox and vaccinia viruses by polymerase chain reaction.pcr following two methods of dna extraction was used to confirm the growth of camel pox virus (cpv) and vaccinia virus in cell culture and chorioallantoic membrane. results were compared with the commonly used neutralization test. the first method of dna extraction was accomplished by using viral dna in tissue culture supernatant and chorioallantoic membrane, which was released by initial heating for 15 min at 99 degrees c followed by ordinary pcr. in the second method dna was extracted by using ...200919484375
patterns of orthopox virus wild rodent hosts in south germany.although cowpox virus (cpxv) infections in a variety of dead-end hosts have been investigated in germany for more than 50 years, data on species and geographical distribution of cpxv in reservoir hosts are sparse. here we present the first comprehensive study of 825 rodents that have been collected in bavaria, southern germany. in summary, six different rodent species (apodemus flavicollis, myodes glareolus, microtus arvalis, apodemus sylvaticus, microtus agrestis, and arvicola amphibius) were t ...200919492947
transgenic chloroplasts are efficient sites for high-yield production of the vaccinia virus envelope protein a27l in plant cellsdagger.orthopoxviruses (opvs) have recently received increasing attention because of their potential use in bioterrorism and the occurrence of zoonotic opv outbreaks, highlighting the need for the development of safe and cost-effective vaccines against smallpox and related viruses. in this respect, the production of subunit protein-based vaccines in transgenic plants is an attractive approach. for this purpose, the a27l immunogenic protein of vaccinia virus was expressed in tobacco using stable transfo ...200919508274
diverse recognition of conserved orthopoxvirus cd8+ t cell epitopes in vaccinated rhesus macaques.vaccinia virus (vacv) induces a vigorous virus-specific cd8+ t cell response that plays an important role in control of poxvirus infection. to identify immunodominant poxvirus proteins and to facilitate future testing of smallpox vaccines in non-human primates, we used an algorithm for the prediction of vacv peptides able to bind to the common macaque mhc class i molecule mamu-a*01. we synthesized 294 peptides derived from 97 vacv orfs; 100 of these peptides did not contain the canonical proline ...200919531389
comparative sequence analysis of a-type inclusion (ati) and p4c proteins of orthopoxviruses that produce typical and atypical ati phenotypes.some orthopoxviruses produce large proteinaceous intracellular bodies, known as a-type inclusions (atis) during infection of host cells. virions associate with atis resulting in distinct phenotypes referred to as v+, v+/ and v⁻. the phenotype v+ has the virions embedded in the ati matrix; v⁻ has no virions embedded within or on the surface of the ati matrix, whereas an aberrant phenotype, the v+/ has virions only on the surface of atis. viruses that do not produce ati are designated as v⁰. recom ...200919533319
the new acam2000 vaccine and other therapies to control orthopoxvirus outbreaks and bioterror attacks.quarantine, case tracing and population vaccination facilitated the global eradication, in 1980, of variola virus, the causative agent of smallpox. the vaccines used during the eradication period, including dryvax, the smallpox vaccine used in the usa, were live vaccinia and cowpoxvirus-based vaccines, which induced long-lasting and cross-protective immunity against variola and other related poxviruses. these vaccine viruses were produced by serial propagation in domesticated animals. the drawba ...200919538111
immunogenicity and protective efficacy of semliki forest virus replicon-based dna vaccines encoding goatpox virus structural proteins.goatpox, caused by goatpox virus (gtpv), is an acute feverish and contagious disease in goats often associated with high morbidity and high mortality. to resolve potential safety risks and vaccination side effects of existing live attenuated goatpox vaccine (av41), two semliki forest virus (sfv) replicon-based bicistronic expression dna vaccines (pcsm-aal and pcsm-baa) which encode gtpv structural proteins corresponding to the vaccinia virus proteins a27, l1, a33, and b5, respectively, were cons ...200919559453
comparison and phylogenetic analysis of cowpox viruses isolated from cats and humans in fennoscandia.cowpox virus (cpxv), a member of the genus orthopoxvirus (opv), has reservoirs in small mammals and may cause disease in humans, felidae and other animals. in this study we compared cpxvs isolated from humans and cats in fennoscandia by restriction enzyme and dna sequence analysis. the hindiii restriction profiles clearly distinguished geographically distinct cpxv isolates, whereas only minor differences were found between the profiles of geographically linked isolates. the complete gene sequenc ...200919585075
evolution of the schlafen genes, a gene family associated with embryonic lethality, meiotic drive, immune processes and orthopoxvirus virulence.genes of the schlafen family, first discovered in mouse, are expressed in hematopoietic cells and are involved in immune processes. previous results showed that they are candidate genes for two major phenomena: meiotic drive and embryonic lethality (ddk syndrome). however, these genes remain poorly understood, mostly due to the limitations imposed by their similarity, close location and the potential functional redundancy of the gene family members. here we use genomic and phylogenetic studies t ...200919619625
rapid and high-throughput pan-orthopoxvirus detection and identification using pcr and mass spectrometry.the genus orthopoxvirus contains several species of related viruses, including the causative agent of smallpox (variola virus). in addition to smallpox, several other members of the genus are capable of causing human infection, including monkeypox, cowpox, and other zoonotic rodent-borne poxviruses. therefore, a single assay that can accurately identify all orthopoxviruses could provide a valuable tool for rapid broad orthopovirus identification. we have developed a pan-orthopoxvirus assay for i ...200919623263
analysis of vaccinia virus-host protein-protein interactions: validations of yeast two-hybrid screenings.vaccinia virus, a large double-stranded dna virus, is the prototype of the orthopoxvirus genus, which includes several pathogenic poxviruses of humans, such as monkeypox virus and variola virus. here, we report a comprehensive yeast two-hybrid (y2h) screening for the protein-protein interactions between vaccinia and human proteins. a total of 109 novel vaccinia-human protein interactions were detected among 33 viral proteins. to validate subsets of those interactions, we constructed an orfeome l ...200919637933
therapeutic and prophylactic drugs to treat orthopoxvirus infections.with the global eradication of smallpox in 1979, the causative agent, variola, no longer circulates in human populations. other human poxvirus infections, such as those caused by vaccinia, cowpox virus and molluscum, are usually relatively benign in immunocompetent individuals. conversely, monkeypox virus infections cause high levels of mortality and morbidity in africa and the virus appears to be increasing its host range, virulence and demographic environs. furthermore, there are concerns that ...200819727418
when good vaccines go wild: feral orthopoxvirus in developing countries and beyond.the presence of zoonotic poxviruses in nature represents a potential human health risk that has to be re-evaluated by health authorities not only in developing countries, but also in many developed countries. for example, buffalopox virus infection remains to be a threat to humans and cattle in india, and monkeypox virus infection persists in several inhabited places in africa and, more recently, in the usa. there are also a great number of zoonotic transmissions of cowpox virus from cats to hum ...200819738346
nested-multiplex pcr detection of orthopoxvirus and parapoxvirus directly from exanthematic clinical samples.orthopoxvirus (opv) and parapoxvirus (ppv) have been associated with worldwide exanthematic outbreaks. some species of these genera are able to infect humans and domestic animals, causing serious economic losses and public health impact. rapid, useful and highly specific methods are required to detect and epidemiologically monitor such poxviruses. in the present paper, we describe the development of a nested-multiplex pcr method for the simultaneous detection of opv and ppv species directly from ...200919747382
st-246 inhibits in vivo poxvirus dissemination, virus shedding, and systemic disease manifestation.orthopoxvirus infections, such as smallpox, can lead to severe systemic disease and result in considerable morbidity and mortality in immunologically naïve individuals. treatment with st-246, a small-molecule inhibitor of virus egress, has been shown to provide protection against severe disease and death induced by several members of the poxvirus family, including vaccinia, variola, and monkeypox viruses. here, we show that st-246 treatment not only results in the significant inhibition of vacci ...200919752270
new classes of orthopoxvirus vaccine candidates by functionally screening a synthetic library for protective antigens.the licensed smallpox vaccine, comprised of infectious vaccinia, is no longer popular as it is associated with a variety of adverse events. safer vaccines have been explored such as further attenuated viruses and component designs. however, these alternatives typically provide compromised breadth and strength of protection. we conducted a genome-level screening of cowpox, the ancestral poxvirus, in the broadly immune-presenting c57bl/6 mouse as an approach to discovering novel components with pr ...200919800089
antiviral evaluation of octadecyloxyethyl esters of (s)-3-hydroxy-2-(phosphonomethoxy)propyl nucleosides against herpesviruses and orthopoxviruses.our previous studies showed that esterification of 9-(s)-[3-hydroxy-2-(phosphonomethoxy)propyl]adenine (hpmpa) or 1-(s)-[3-hydroxy-2-(phosphonomethoxy)-propyl]cytosine (hpmpc) with alkoxyalkyl groups such as hexadecyloxypropyl (hdp) or octadecyloxyethyl (ode) resulted in large increases in antiviral activity and oral bioavailability. the hdp and ode esters of hpmpa were shown to be active in cells infected with human immunodeficiency virus, type 1 (hiv-1), while hpmpa itself was virtually inacti ...200919800369
[orthopoxvirus genes for kelch-like proteins. iii. construction of mousepox (ectromelia) virus variants with targeted gene deletions].mousepox (ectromelia) virus genome contains four genes encoding for kelch-like proteins evm018, evm027, evm150 and evm167. a complete set of insertion plasmids was constructed to allow the production of recombinant ectromelia viruses with targeted deletions of one to four genes of kelch family both individually (single mutants) and in different combinations (double, triple and quadruple mutants). it was shown that deletion of any of the three genes evmo18, evm027 or evm167 resulted in reduction ...200919807023
molecular smallpox vaccine delivered by alphavirus replicons elicits protective immunity in mice and non-human primates.naturally occurring smallpox was eradicated as a result of successful vaccination campaigns during the 1960s and 1970s. because of its highly contagious nature and high mortality rate, smallpox has significant potential as a biological weapon. unfortunately, the current vaccine for orthopoxviruses is contraindicated for large portions of the population. thus, there is a need for new, safe, and effective orthopoxvirus vaccines. alphavirus replicon vectors, derived from strains of venezuelan equin ...200919833247
the phylogenetics and ecology of the orthopoxviruses endemic to north america.the data presented herein support the north american orthopoxviruses (na opxv) in a sister relationship to all other currently described orthopoxvirus (opxv) species. this phylogenetic analysis reaffirms the identification of the na opxv as close relatives of "old world" (eurasian and african) opxv and presents high support for deeper nodes within the chordopoxvirinae family. the natural reservoir host(s) for many of the described opxv species remains unknown although a clear virus-host associat ...200919865479
long-lasting immunity by early infection of maternal-antibody-protected infants.newborn higher vertebrates are largely immuno-incompetent and generally survive infections--including poxviruses--by maternal antibody protection. here, we show that mice survived epidemics as adults only if exposed to lethal orthopoxvirus infections during infancy under the umbrella of maternal protective antibodies. this implies that both the absence of exposure to infection during early infancy or of effective vaccination renders the population highly susceptible to new or old re-emerging pat ...201019877011
tissue damage caused by animal orthopoxviruses cowpox, ectromelia, vaccinia and parapoxvirus ovis in human cornea. 201019878105
multicolour, multiplex real-time pcr assay for the detection of human-pathogenic poxviruses.after eradication of variola virus, one of the most dangerous infectious diseases affecting mankind, today other poxviruses of different genera can cause infection in humans. these viruses include human-specific molluscipoxviruses as well as zoonotic orthopoxviruses and parapoxviruses. while non-variola orthopoxvirus infections mostly cause mild symptoms in immunocompetent persons, they can evoke severe disease in immunocompromised patients. since the typical poxviral skin lesions are rarely dia ...201019879351
how long ago did smallpox virus emerge?unlike vertebrates, for which paleontological data are available, and rna viruses, which display a high rate of genetic variation, an objective estimate of time parameters for the molecular evolution of dna viruses, which display a low rate of accumulation of mutations, is a complex problem. genomic studies of a set of smallpox (variola) virus (varv) isolates demonstrated the patterns of phylogenetic relationships between geographic variants of this virus. using archival data on smallpox outbrea ...200919882103
in vitro prevention of vaccinia and herpesvirus infection spread in explanted human corneas by n-chlorotaurine.weak oxidants produced by activated human leukocytes are proven antimicrobial substances. we tested whether n-chlorotaurine (nct, taurine chloramine), the chlorinated metabolite of the amino acid taurine, in addition to direct virucidal effects on viral suspensions, has the capability to prevent cell-to-cell spread of viruses in human corneal epithelium.201019887880
inhibition of monkeypox virus replication by rna interference.the orthopoxvirus genus of poxviridae family is comprised of several human pathogens, including cowpox (cpxv), vaccinia (vacv), monkeypox (mpv) and variola (varv) viruses. species of this virus genus cause human diseases with various severities and outcome ranging from mild conditions to death in fulminating cases. currently, vaccination is the only protective measure against infection with these viruses and no licensed antiviral drug therapy is available. in this study, we investigated the pote ...200919889227
vaccinia virus a43r gene encodes an orthopoxvirus-specific late non-virion type-1 membrane protein that is dispensable for replication but enhances intradermal lesion formation.the vaccinia virus a43r open reading frame encodes a 168-amino acid protein with a predicted n-terminal signal sequence and a c-terminal transmembrane domain. although a43r is conserved in all sequenced members of the orthopoxvirus genus, no non-orthopoxvirus homolog or functional motif was recognized. biochemical and confocal microscopic studies indicated that a43 is expressed at late times following viral dna synthesis and is a type-1 membrane protein with two n-linked oligosaccharide chains. ...201019900687
nck- and n-wasp-dependent actin-based motility is conserved in divergent vertebrate poxviruses.vaccinia virus enhances its cell-to-cell spread by stimulating actin polymerization via src- and abl-mediated phosphorylation of the highly conserved orthopoxvirus protein a36. the yatapoxvirus, yaba-like disease virus (yldv), also induces actin polymerization, although it lacks an obvious a36 ortholog. we found that the yldv protein yl126 can functionally replace a36 to promote nck- and n-wasp-dependent actin polymerization. at least five phosphorylated tyrosines in yl126, rather than a single ...200920006842
studying nk cell responses to ectromelia virus infections in mice.here we describe methods for the in vivo study of antiviral nk cell responses using the mouse orthopoxvirus ectromelia virus as a model, the agent of mousepox. the methods include those specific for the preparation and use of ectromelia virus such as the production of virus stocks in tissue culture and in live mice, the purification of virus stocks, the titration of virus stocks and virus loads in organs, and the infection of mice. the chapter also includes methods for the specific study of nk c ...201020033657
inhibition of type iii interferon activity by orthopoxvirus immunomodulatory proteins.the type iii interferon (ifn) family elicits an antiviral response that is nearly identical to that evoked by ifn-alpha/beta. however, these cytokines (known as ifn-lambda1, 2, and 3) signal through a distinct receptor, and thus may be resistant to the evasion strategies used by some viruses to avoid the ifn-alpha/beta response. orthopoxviruses are highly resistant to ifn-alpha/beta because they encode well-characterized immunomodulatory proteins that inhibit ifn activity. these include a secret ...201020038204
efficacy of st-246 versus lethal poxvirus challenge in immunodeficient mice.the threat of smallpox as a bioweapon and the emerging threat of human monkeypox, among other poxviral diseases, highlight the need for effective poxvirus countermeasures. st-246, which targets the f13l protein in vaccinia virus and its homologs in other orthopoxvirus species, provides full protection from lethal poxviral disease in numerous animal models and seems to be safe in humans. all previous evaluations of st-246 efficacy have been in immunocompetent animals. however, the risk of severe ...201020080762
epidermal injury and infection during poxvirus immunization is crucial for the generation of highly protective t cell-mediated immunity.variola major (smallpox) infection claimed hundreds of millions lives before it was eradicated by a simple vaccination strategy: epicutaneous application of the related orthopoxvirus vaccinia virus (vacv) to superficially injured skin (skin scarification, s.s.). however, the remarkable success of this strategy was attributed to the immunogenicity of vacv rather than to the unique mode of vaccine delivery. we now show that vacv immunization via s.s., but not conventional injection routes, is esse ...201020081864
an outbreak of orthopoxvirus-associated disease in horses in southern brazil.an outbreak of severe cutaneous disease associated with an orthopoxvirus in horses in southern brazil is described. fourteen crioulo mares and foals from a husbandry farm developed papules, and vesicles progressing to proliferative and exudative lesions on the muzzle, external nares, and external and internal lips. the vesicles eroded, and the proliferative lesions eventually bled and progressed to moist crusts and scars. the clinical signs lasted approximately 6-12 days, after which the animals ...201020093706
comparison of west african and congo basin monkeypox viruses in balb/c and c57bl/6 mice.although monkeypox virus (mpxv) studies in wild rodents and non-human primates have generated important knowledge regarding mpxv pathogenesis and inferences about disease transmission, it might be easier to dissect the importance of virulence factors and correlates of protection to mpxv in an inbred mouse model. herein, we compared the two clades of mpxv via two routes of infection in the balb/c and c57bl/6 inbred mice strains. our studies show that similar to previous animal studies, the congo ...201020111702
a mouse model of lethal infection for evaluating prophylactics and therapeutics against monkeypox virus.monkeypox virus (mpxv) is an orthopoxvirus closely related to variola, the etiological agent of smallpox. in humans, mpxv causes a disease similar to smallpox and is considered to be an emerging infectious disease. moreover, the use of mpxv for bioterroristic/biowarfare activities is of significant concern. available small animal models of human monkeypox have been restricted to mammals with poorly defined biologies that also have limited reagent availability. we have established a murine mpxv m ...201020130052
application of the ibis-t5000 pan-orthopoxvirus assay to quantitatively detect monkeypox viral loads in clinical specimens from macaques experimentally infected with aerosolized monkeypox virus.monkeypox virus (mpxv), a member of the family poxviridae and genus orthopoxvirus, causes a smallpox-like disease in humans. a previously described pan-orthopoxvirus assay, based on a broad-range polymerase chain reaction (pcr) coupled with electrospray ionization mass spectrometry (pcr/esi-ms), was evaluated for its ability to detect mpxv from spiked human and aerosol-infected cynomolgous macaque (macaca fascicularis) samples. detection of mpxv dna from macaque tissue, blood, and spiked human b ...201020134011
potent oncolytic activity of raccoonpox virus in the absence of natural pathogenicity.a number of oncolytic virus (ov) candidates currently in clinical trials are human viruses that have been engineered to be safer for patient administration by limiting normal cell targeting and replication. the newest ovs include viruses that cause no disease in humans, yet still have natural tumor tropism. raccoonpox virus (rcnv) is a member of the orthopoxvirus genus of poxviridae and closely related to vaccinia virus, yet has no known pathogenicity in any mammalian species. a screen of cells ...201020160706
rapid detection of orthopoxvirus by semi-nested pcr directly from clinical specimens: a useful alternative for routine laboratories.orthopoxvirus (opv) has been associated with worldwide exanthematic outbreaks, which have resulted in serious economic losses as well as impact on public health. although the current classical and molecular methods are useful for the diagnosis of opv, they are largely inaccessible to unsophisticated clinical laboratories. the major reason for the inaccessibility is that they require both virus isolation and dna manipulation. in this report, a rapid, sensitive and low-cost semi-nested pcr method ...201020166167
tecovirimat, a p37 envelope protein inhibitor for the treatment of smallpox infection.since the eradication of naturally occurring smallpox in 1980, the fear that variola virus could be used as a biological weapon has become real. over the last 10 years, emergency preparedness programs have been launched to protect populations against a smallpox outbreak or the possible emergence in humans of other orthopoxvirus infections, such as monkeypox. vaccination against smallpox was responsible for its eradication, but was linked with high rates of adverse events and contraindications. i ...201020191435
[human infections caused by vaccinia-like poxviruses in brazil].since 1999, human infection caused by orthopoxvirus has been observed in at least eight brazilian states, with the presence of vesicles that evolve to pustules and crusts, especially on the hands, arms and face, after contact with cows showing comparable lesions on the udder. in addition to the skin lesions, there have been descriptions of patients with axillary ganglionic reactions that are sometimes painful, along with fever, headache, fatigue, dehydration, anorexia, sudoresis, arthralgia and ...200920209353
a silent enzootic of an orthopoxvirus in ghana, west africa: evidence for multi-species involvement in the absence of widespread human disease.human monkeypox has never been reported in ghana, but rodents captured in forested areas of southern ghana were the source of the monkeypox virus introduced into the united states in 2003. subsequent to the outbreak in the united states, 204 animals were collected from two commercial trapping sites in ghana. animal tissues were examined for the presence of orthopoxvirus (opxv) dna using a real-time polymerase chain reaction, and sera were assayed for antibodies against opxv. animals from five ge ...201020348530
isolation and characterization of indian isolates of camel pox virus.in this study, we isolated and identified three camel pox viruses (cmlv) from two outbreaks of camel pox infection in camels associated with eruptions on cheeks, nostrils, limbs, scrotum, and sheath that occurred at different places of bikaner district, rajasthan (india). the scab specimens collected were subjected for virus isolation in vero cell culture, and the isolated viruses were characterized by employing polymerase chain reaction (pcr) and sequencing. the causative agent was identified a ...201020358400
dosage comparison of congo basin and west african strains of monkeypox virus using a prairie dog animal model of systemic orthopoxvirus disease.the prairie dog is valuable for the study of monkeypox virus (mpxv) virulence and closely resembles human systemic orthopoxvirus disease. herein, we utilize a variable dose intranasal challenge with approximately 10(3), 10(4), 10(5), and 10(6)pfu for each clade to further characterize virulence differences between the two mpxv clades. a trend of increased morbidity and mortality as well as greater viral shedding was observed with increasing viral challenge dose. additionally, there appeared to b ...201020374968
safety and pharmacokinetics of the antiorthopoxvirus compound st-246 following repeat oral dosing in healthy adult subjects.st-246, a novel compound that inhibits egress of orthopoxvirus from infected cells, is being evaluated as a treatment for pathogenic orthopoxvirus infections in humans. this phase i, double-blind, randomized, placebo-controlled, escalating multiple-dose study was conducted to determine the safety, tolerability, and pharmacokinetics of st-246 administered as a single daily oral dose of 250, 400, or 800 mg for 21 days to nonfasting healthy human volunteers. st-246 appeared to be well tolerated, wi ...201020385870
seroprevalence of orthopoxvirus in an amazonian rural village, acre, brazil.vaccinia virus strains from the family poxviridae have been frequently isolated in brazil and associated with outbreaks of exanthematic disease affecting cows and humans. an elisa igg was applied to evaluate the seroprevalence of orthopoxviruses in a community located in a rural settlement in the amazon region, where no orthopoxvirus outbreaks have yet been reported. an overall seroprevalence of 27.89% was found, and it was 23.38% in the non-vaccinated population (smallpox vaccination). these re ...201020428903
a novel highly reproducible and lethal nonhuman primate model for orthopox virus infection.the intentional re-introduction of variola virus (varv), the agent of smallpox, into the human population is of great concern due its bio-terroristic potential. moreover, zoonotic infections with cowpox (cpxv) and monkeypox virus (mpxv) cause severe diseases in humans. smallpox vaccines presently available can have severe adverse effects that are no longer acceptable. the efficacy and safety of new vaccines and antiviral drugs for use in humans can only be demonstrated in animal models. the exis ...201020454688
fatal cowpox virus infection in captive banded mongooses (mungos mungo).cowpox virus infections have been described in various domestic and exotic animal species. this report is the first on an outbreak of fatal generalized cowpox virus infection among captive banded mongooses (mungos mungo, suborder feliformia). all animals of a colony of 8 mongooses showed a fulminant course of disease. the whole population died (n=7) or was euthanized (n=1) within 11 days. postmortem examinations were performed on 4 animals. all animals showed extensive necrotizing inflammation o ...201020472807
congregation of orthopoxvirus virions in cytoplasmic a-type inclusions is mediated by interactions of a bridging protein (a26p) with a matrix protein (atip) and a virion membrane-associated protein (a27p).some orthopoxviruses, e.g., the cowpox, ectromelia, and raccoonpox viruses, form large, discrete cytoplasmic inclusions within which mature virions (mvs) are embedded by a process called occlusion. these inclusions, which may protect occluded mvs in the environment, are composed of aggregates of the a-type inclusion protein (atip), which is truncated in orthopoxviruses such as vaccinia virus (vacv) and variola virus that fail to form inclusions. in addition to an intact atip, occlusion requires ...201020484506
vaccinia virus infection in monkeys, brazilian amazon.to detect orthopoxvirus in the brazilian amazon, we conducted a serosurvey of 344 wild animals. neutralizing antibodies against orthopoxvirus were detected by plaque-reduction neutralizing tests in 84 serum samples. amplicons from 6 monkey samples were sequenced. these amplicons identified vaccinia virus genetically similar to strains from bovine vaccinia outbreaks in brazil.201020507750
comparative whole genome sequence analysis of wild-type and cidofovir-resistant monkeypoxvirus.we performed whole genome sequencing of a cidofovir {[(s)-1-(3-hydroxy-2-phosphonylmethoxy-propyl) cytosine] [hpmpc]}-resistant (cdv-r) strain of monkeypoxvirus (mpv). whole-genome comparison with the wild-type (wt) strain revealed 55 single-nucleotide polymorphisms (snps) and one tandem-repeat contraction. over one-third of all identified snps were located within genes comprising the poxvirus replication complex, including the dna polymerase, rna polymerase, mrna capping methyltransferase, dna ...201020509894
cutting edge: fcr-like 5 on innate b cells is targeted by a poxvirus mhc class i-like immunoevasin.under selective pressure from host immunity, viruses have retained genes encoding immunoevasins, molecules interfering with host viral recognition and clearance. due to their binding specificities, immunoevasins can be exploited as affinity labels to identify host-encoded molecules of previously unsuspected importance in defense against the relevant class of virus. we previously described an orthopoxvirus mhc class i-like protein (omcp) that binds with high affinity to the activating receptor nk ...201020519648
physiologic reference ranges for captive black-tailed prairie dogs (cynomys ludovicianus).the black-tailed prairie dog (cynomys ludovicianus) is a member of the order rodentia and the family sciuridae. ecologically, prairie dogs are a keystone species in prairie ecology. this species is used as an animal model for human gallbladder disease and diseases caused by infection with clostridium difficile, yersinia pestis, francisella tularensis, and most recently, orthopoxvirus. despite increasing numbers of prairie dogs used in research and kept as pets, few data are available on their ba ...201020587156
development of real-time pcr assay for specific detection of cowpox virus.the number of recorded human cowpox cases are recently increasing. the symptoms caused by cowpox virus (cpxv) in a number of human cases are close to the symptoms characteristic of the orthopoxviral human infections caused by monkeypox or smallpox (variola) viruses. any rapid and reliable real-time pcr method for distinguishing cowpox from smallpox and monkeypox is yet absent.201020594906
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