| isolation and serial propagation of porcine epidemic diarrhea virus in cell cultures and partial characterization of the isolate. | porcine epidemic diarrhea virus (pedv) was isolated in vero cell cultures from the small intestine of a piglet experimentally infected with porcine coronavirus 83p-5, that had been isolated during outbreaks of porcine acute diarrhea and passaged in piglets. the isolation of the pedv was successful only in vero cells maintained in the maintenance medium (mm) containing trypsin. infected vero cell cultures exhibited cpe characterized by cell-fusion and syncytial formation, as well as cytoplasmic f ... | 1992 | 1318752 |
| coronavirus infection in mink (mustela vison). serological evidence of infection with a coronavirus related to transmissible gastroenteritis virus and porcine epidemic diarrhea virus. | antibodies to a transmissible gastroenteritis virus (tgev)-related coronavirus have been demonstrated in mink sera by indirect immunofluorescence, peroxidase-linked antibody assays and immunoblotting. this is the first serological evidence of a specific coronavirus infection in mink. the putative mink coronavirus (mcv) seems to be widespread in the danish mink population with a prevalence approaching 100%. analysis by immunoblotting has shown that mcv is closely related to tgev by the spike (s), ... | 1992 | 1319622 |
| an elisa for detection of antibodies against porcine epidemic diarrhoea virus (pedv) based on the specific solubility of the viral surface glycoprotein. | viral proteins of porcine epidemic diarrhoea virus (pedv) were extracted from the cytoplasm of infected vero cells using hypotonic conditions and a non-ionic detergent. both the ph and the nacl concentration of the extraction buffer were varied in attempts to increase the solubility of the virion spike glycoproteins (s-protein) and of the nucleocapsid proteins (n-protein). monoclonal antibodies, hyperimmune sera and convalescent pig sera were used to identify and monitor these proteins by immuno ... | 1992 | 1441196 |
| use of two enzyme-linked immunosorbent assays to monitor antibody responses in swine with experimentally induced infection with porcine epidemic diarrhea virus. | a blocking elisa was developed to detect antibodies directed against porcine epidemic diarrhea virus (pedv). the pedv antigen was first incubated with dilutions of test sera. any antigen that was not blocked by antibodies in the serum was assayed in a double-antibody sandwich elisa, using 2 monoclonal antibodies directed against different antigenic sites on pedv as capture and detecting antibodies, respectively. the blocking elisa was compared with a fixed-cell elisa that used monolayers of vero ... | 1991 | 1654032 |
| quantitation, biological and physicochemical properties of cell culture-adapted porcine epidemic diarrhea coronavirus (pedv). | the porcine epidemic coronavirus (pedv), tentatively classified as a coronavirus, was adapted to vero cells and a plaque test developed for infectivity titration, allowing us to test the biological and biophysical properties of the virus. growth kinetics showed peak titers of 10(5.5) plaque-forming units ml-1 15 h after infection. filtration experiments and electron microscopy revealed a particle diameter between 100 and 200 nm. the buoyant density of the virus was 1.18. the particle lost its in ... | 1989 | 2549681 |
| propagation of the virus of porcine epidemic diarrhea in cell culture. | porcine epidemic diarrhea virus (pedv) was adapted to serial propagation in vero cell cultures by adding trypsin to the medium. pedv-infected cells showed a distinct cytoplasmic fluorescence when examined by a fluorescent-antibody-staining technique. cytopathic effects, such as vacuolation, formation of syncytia, and fusion of cells, were detected even at passage 1 of the pedv in vero cells. once adapted, the virus induced numerous syncytia containing over 100 nuclei. from virus passage 5 on, al ... | 1988 | 2853174 |
| comparison of two methods for detection of transmissible gastroenteritis virus in feces of pigs with experimentally induced infection. | an indirect, double-antibody sandwich-type elisa for detection of transmissible gastroenteritis virus (tgev) was developed, using a solid phase of rabbit hyperimmune serum and a pool of 3 antipeplomer monoclonal antibodies to trap and to detect the virus, respectively. the technique was used to detect viral antigen in feces of pigs that had been infected with the virulent miller strain, the attenuated purdue strain, or the erica strain (a dutch field isolate) of tgev. the results were compared w ... | 1988 | 2854707 |
| pathogenicity and persistence of pleural effusion disease virus isolates in rabbits. | nine isolates of pleural effusion disease agent or virus (pedv) from treponema-infected rabbits in various countries were examined for pathogenicity and persistence in rabbits. the isolates showed a wide range of pathogenicity and were categorized into three groups according to the severity of the acute infection. group 1 comprised isolates causing more than 50% mortality, group 2 isolates causing mortality below 50%, while group 3 comprised isolates causing almost subclinical infections. the ra ... | 1986 | 3086501 |
| porcine epidemic diarrhea virus (cv 777) and feline infectious peritonitis virus (fipv) are antigenically related. | using gut sections from pigs infected with porcine epidemic diarrhea virus (strain cv 777) and ascitic fluid from cats which had succumbed to feline infectious peritonitis (fip), a weak cross reaction was found by immunofluorescence. its specificity was confirmed when detergent-treated purified cv 777 showed a prominent reaction with fipv antibodies in elisa; no reaction was obtained with intact virions, which indicated common determinants on an internal component of the particle. antigenic cros ... | 1988 | 3196169 |
| pathogenetic observations on pleural effusion disease in rabbits. | a pathogenetic study of pleural effusion disease (ped) in rabbits was made, using the virulent ped agent or virus (pedv) and an avirulent derivate of this isolate. independent of infective dose within the range examined, the virulent isolate caused fatal clinical disease, whereas the avirulent isolate caused subclinical infection. the two isolates differed in rapidity of initial spread of infection and in the maximum virus titres in serum, but they both resulted in a similar low level persisting ... | 1985 | 3994516 |
| significance of lysosomes in the morphogenesis of coronaviruses. | virus-containing electron-dense membrane-bound cytoplasmic bodies are described in tracheal epithelial cells of chickens infected with infectious bronchitis virus and in intestinal epithelial cells of swine infected with porcine epidemic diarrhea virus. using silver-methenamine staining, phosphotungstic acid staining and acid phosphatase enzyme cytochemical staining of ultra-thin sections, these bodies were shown to be virus-containing secondary lysosomes and residual bodies. the accumulation of ... | 1984 | 6320768 |
| pleural effusion disease in rabbits. properties of the aetiological agent. | the size and heat sensitivity of pleural effusion disease (ped) agent or virus (pedv) propagated in rabbits were examined. the infectious particles were estimated to be between 25 and 50 nm by filtration. residual infectivity of infectious serum was 0.1 per cent after heating at 56 degrees c for 4 hours. pedv and the stockholm agent appeared identical concerning pathogenic and immunogenic properties by infection experiments and protection tests in rabbits. two of the three pedv isolates were les ... | 1983 | 6409056 |
| pleural effusion disease in rabbits. observations on viraemia, immunity and transmissibility. | baby rabbits surviving infection with pleural effusion disease virus (pedv) developed viraemia persisting for at least six months. only the infectious serum samples collected during the first 2 months of disease could transfer the typical ped. six months after neonatal infection, virus concentration in serum was 102 to 104 rabbit-infectious doses per ml, the level of igg appeared elevated, and serum rendered non-infectious by ether-treatment had a protective effect in passive immunisation experi ... | 1981 | 7332487 |
| evaluation of a blocking elisa using monoclonal antibodies for the detection of porcine epidemic diarrhea virus and its antibodies. | two blocking enzyme-linked immunosorbent assays (elisas) involving the use of monoclonal antibodies (mab) as capture and detecting agents and an indirect fluorescence test (ift) were used for the detection of porcine epidemic diarrhea virus (pedv) antigen or its antibodies. in the elisa for viral antigen detection, the blocking step is accomplished by incubation of fecal samples, in duplicate wells, with pedv-specific positive or negative serum, whereas in the elisa for antibody detection the bl ... | 1995 | 7779966 |
| prevalence of infections with enzootic respiratory and enteric viruses in feeder pigs entering fattening herds. | the prevalence of infections with h1n1- and h3n2-influenza viruses, porcine respiratory coronavirus (prcv), transmissible gastroenteritis virus (tgev) and porcine epidemic diarrhoea virus (pedv) in feeder pigs shortly after their entry into fattening units was examined. ten groups of pigs with acute respiratory disease during the months september to october 1991 and seven groups of pigs with acute diarrhoea during the months february to march 1992 were investigated. on arrival in the fattening h ... | 1994 | 7900243 |
| sequence of the spike protein of the porcine epidemic diarrhoea virus. | the complete sequence of the spike (s) gene of the br1/87 isolate of porcine epidemic diarrhoea virus (pedv) was determined from cdna clones. the predicted polypeptide was 1383 amino acids long, contained 29 potential n-linked glycosylation sites and showed structural features similar to those of the coronavirus spike protein. the pedv s protein, like that of the members of the transmissible gastroenteritis virus (tgev)-related subset, lacks a proteolytic site to yield cleaved amino and carboxy ... | 1994 | 8176382 |
| preliminary characterization of a monoclonal antibody specific for a viral 27 kd glycoprotein family synthesized in porcine epidemic diarrhoea virus infected cells. | we describe a new monoclonal antibody no. 204 (mcab 204) which recognized a family of four polypeptides, consisting of a 27kd, a 24/23kd double band and a 19kd protein present within pedv infected cell lysates. these proteins were identified by immunoprecipitation as well as by staining of immunoblots. in infected vero cell cultures, the synthesis of the 27kd protein was initiated between 6 and 8 hours post inoculation. the 24/23kd double band and the 19kd protein were only detectable later. at ... | 1993 | 8209730 |
| sequence analysis of the nucleocapsid protein gene of porcine epidemic diarrhoea virus. | the nucleotide (nt) sequence of 1.7 kbp cdna representing the 3' end of the pedv genome has been determined. viral rna was reverse transcribed and the cdna was amplified by polymerase chain reaction using degenerate primers. the sequences of the primers were based on conserved regions of coronaviral genomes. a 1323 nt open reading frame (orf) showed good homology to the nucleocapsid (n) gene of other coronaviruses. the greatest homologies at the amino acid and the nt levels were observed with hu ... | 1993 | 8209770 |
| genome organization of porcine epidemic diarrhoea virus. | in order to study the organization of the genome of porcine epidemic diarrhoea virus (pedv), we constructed a cdna library in a phage expression vector by using poly(a) rna from pedv-infected vero cells. an anti-pedv hyperimmune serum was used to probe the library. the first isolated clone mapped within the n gene and was subsequently used for rescreening the library. the selected clones allowed us to establish the sequence of the 3'-most 7.4 kb of the pedv genome. analysis of the cdna sequences ... | 1993 | 8209771 |
| sequence analysis of the porcine epidemic diarrhea virus genome between the nucleocapsid and spike protein genes reveals a polymorphic orf. | in order to investigate the genome organization of the porcine epidemic diarrhea virus (pedv) further, cdna clones covering the region between the nucleocapsid and the spike (s) protein genes were independently constructed and sequenced for the two virulent isolates br1/87 and cv777. of the three major orfs identified, two were found to encode the major and minor coronavirus membrane proteins m and sm. a potentially single orf, designated orf3 according to the pattern of the viral subgenomic mrn ... | 1994 | 8291230 |
| porcine epidemic diarrhoea virus as a cause of persistent diarrhoea in a herd of breeding and finishing pigs. | a clinical and virological study of an outbreak of porcine epidemic diarrhoea in a combined breeding and finishing pig herd used elisa techniques to detect porcine epidemic diarrhoea virus in faeces and antibodies in blood. no seropositive pigs were found at the start of the outbreak. the first signs of the disease were observed in fattening pigs and the infection spread rapidly to pregnant sows, farrowing sows and their suckling pigs, gilts and weaners housed in separate barns. depression and d ... | 1993 | 8383370 |
| [detection of porcine epidemic diarrhea virus using electron microscopy in the czech republic]. | coronavirus-induced porcine epidemic diarrhoea (ped) was diagnosed in two swine herds. the causal agent was demonstrated in intestinal contents by electron microscopy and identified by immunoelectron microscopy using specific immune serum to the reference strain ped-cv77. experimental transmission to hysterectomy-derived, colostrum-deprived piglets with an intestinal contents filtrate was successful. the virus was demonstrable by electron microscopy in the intestinal contents between 12th hour a ... | 1993 | 8394039 |
| sequence determination of the nucleocapsid protein gene of the porcine epidemic diarrhoea virus confirms that this virus is a coronavirus related to human coronavirus 229e and porcine transmissible gastroenteritis virus. | the nucleotide sequence of 1.7 kbp cdna, comprising the region nearest the 3' end of the genome of the porcine epidemic diarrhoea virus (pedv), has been independently determined for two european isolates of pedv. almost identical results were obtained for the two isolates, which were derived from cases of pedv infection in belgium and britain in 1977 and 1987, respectively. the sequences contained a 1323 nucleotide (nt) open reading frame (orf), which showed moderate identity to the nucleocapsid ... | 1993 | 8397280 |
| [identification and characterization of new and unknown coronaviruses using rt-pcr and degenerate primers]. | a modified method of the reverse transcription followed by polymerase chain reaction (rt-pcr) was developed in order to examine the genome of a recently discovered virus, the porcine epidemic diarrhoea virus (pedv), which resembled morphologically the coronaviruses. the published sequences of the genomes of various coronaviruses were compared. on the level of the amino acid sequence, conserved regions, common to all coronaviruses, were found in the gene encoding the nonstructural protein 1b as w ... | 1996 | 8720732 |
| enterotoxigenic escherichia coli, rotavirus, porcine epidemic diarrhoea virus, adenovirus and calici-like virus in porcine postweaning diarrhoea in hungary. | in order to obtain data about the significance of enterotoxigenic e. coli (etec), and of the different enteric viruses in the aetiology of porcine postweaning diarrhoea, bacteriological, electron microscopic and elisa studies were made on dead and live pigs. e. coli from the small intestine of diarrhoeal weaned pigs that died were tested for serogroups, pili (fimbriae) and toxin-geno-type. the 108 haemolytic e. coli representing 14 farms and 42 pigs were typed as follows: 0149:k88+etec (56.5%), ... | 1996 | 8826697 |
| studies on the in vitro and in vivo host range of porcine epidemic diarrhoea virus. | | 1995 | 8830468 |
| identification of proteins specified by porcine epidemic diarrhoea virus. | up to now, little was known about the proteins of porcine epidemic diarrhoea virus (pedv). using (i) metabolic labelling, (ii) antisera directed against synthetic peptides and monoclonal antibodies, and (iii) eukaryotic and prokaryotic expression systems, we have started to identify and characterize these proteins. the nucleocapsid protein (n) of pedv was identified as a phosphoprotein with a relative mobility (m(r)) of 57 k. no additional phosphoproteins were detected. at least three glycoprote ... | 1995 | 8830494 |
| rapid diagnosis of porcine epidemic diarrhea virus infection by polymerase chain reaction. | the diagnosis of porcine epidemic diarrhea virus (pedv) infection in the laboratory is rather fastidious because of difficulties in virus propagation. the feasibility of virus propagation in vivo is also limited by the handling of a number of samples at the same time. in this study, the detection of pedv by reverse transcription polymerase chain reaction (rt-pcr) is described. the rt-pcr could detect up to 10(4) tcid50/ml of pedv and did not show any cross reaction with transmissible gastroenter ... | 1997 | 9101487 |
| direct and rapid detection of porcine epidemic diarrhea virus by rt-pcr. | to establish a practical method for detecting porcine epidemic diarrhea virus (pedv), the use of primers derived from sequences that amplify the m protein genes of pedv in a rt-pcr detection system was investigated. primers were designed to amplify a 854-bp fragment by rt-pcr. this reaction was specific to the pedv rna but not to that of other viral genera tested. in experiments with mixtures of pedv and either small intestine or fecal homogenates, this method could detect efficiently the pedv r ... | 1997 | 9504764 |
| immunohistochemical detection of porcine epidemic diarrhea virus compared to other methods. | an immunohistochemistry method using formalin-fixed tissues, a direct immunofluorescence method using cryostat sections, an enzyme-linked immunosorbent assay (elisa), and a pcr method were compared for diagnosis in a litter of weaned pigs that had been experimentally inoculated with wild-type porcine epidemic diarrhea virus (pedv) and killed between 6 and 60 h after onset of diarrhea. the immunohistochemistry method proved to be as reliable as direct immunofluorescence for diagnosis of pedv in t ... | 1998 | 9606002 |
| prokaryotic expression of porcine epidemic diarrhoea virus orf3. | wild type (wt) and cell culture adapted (ca) strains of the coronavirus pedv differ in their ability to cause diarrhea in neonate piglets: the wt strains are virulent; the ca strains are attenuated. comparison of the available nucleotide sequences obtained from the different viral isolates revealed almost complete sequence identity with the exception of variations and truncations in open reading frame 3 (orf3) observed exclusively in ca-pedv isolates. in order to study the biological function(s) ... | 1998 | 9782357 |
| further analysis of the genome of porcine epidemic diarrhoea virus. | we report here the continued determination and analysis of the nucleotide sequence of both wild type (wt) and cell culture adapted (ca) porcine epidemic diarrhoea coronavirus (pedv). these studies were undertaken with two objectives in mind: the identification of common and divergent features in the genomic sequences of wt and ca pedv which can explain the differences in virulence of these isolates and the further exploration of the relationship of pedv to other coronaviruses. | 1998 | 9782358 |
| derivation of attenuated porcine epidemic diarrhea virus (pedv) as vaccine candidate. | the field isolate of porcine epidemic diarrhea virus (pedv) was serially passaged in vero cells. the cell passaged pedv, designated kpedv-9, was tested for its pathogenicity in the neonatal pigs, immunogenicity and safety in the pregnant sows. the result indicated that kpedv-9 at the 93rd passage revealed reduced pathogenicity in the neonatal pigs. pregnant sows inoculated with the attenuated virus showed increased immune responses by elisa. in addition, delivered piglets were protected from cha ... | 1999 | 10418901 |
| detection of porcine epidemic diarrhea virus using polymerase chain reaction and comparison of the nucleocapsid protein genes among strains of the virus. | two pairs of pcr primers were designed to perform nested pcr targetting of a 540 bp fragment of the nucleocapsid (n) protein gene (n gene) of porcine epidemic diarrhea virus (pedv). the n gene of pedv was amplified with 4 pedv strains and 11 small intestines of pedv-infected piglets collected from 2 farms in kagoshima prefecture, japan. nucleotide sequences of the pcr products from a korean and two japanese strains (kkn96-1 and s1) of pedv isolated in 1993 and 1996, respectively, were almost ide ... | 1999 | 10458108 |
| a novel internal open reading frame product expressed from a polycistronic mrna of porcine epidemic diarrhoea virus may not contribute to virus attenuation. | cell-culture-adapted (ca) porcine epidemic diarrhoea virus (pedv) contains three internal open reading frames (i orf) within the nucleocapsid protein gene and lacks the downstream counterpart of porcine transmissible gastroenteritis virus orf7 or feline infectious peritonitis virus orf6a. to confirm whether such features also exist in wild-type (wt) pedv, the 3' 1800 nucleotides of its genome were sequenced and were found to be identical to those of ca virus. the coding potential of i-1 orf was ... | 1999 | 10466791 |
| in situ hybridization for the detection and localization of porcine epidemic diarrhea virus in the intestinal tissues from naturally infected piglets. | detection and localization of porcine epidemic diarrhea virus (pedv) was studied by in situ hybridization with a nonradioactive digoxigenin-labeled probe in formalin-fixed, paraffin-embedded tissues from 10 naturally infected piglets. a 377-base pair cdna probe for viral rna encoding the membrane proteins of pedv cell-culture-adapted strain v215/78 was generated by the reverse transcription polymerase chain reaction. in the retrospective study of pigs from herds with diarrhea, the 10 piglets nat ... | 2000 | 10643982 |
| isolation of porcine epidemic diarrhea virus in porcine cell cultures and experimental infection of pigs of different ages. | this paper describes the isolation of porcine epidemic diarrhea (ped) virus in vero and porcine cell cultures, and the influence of age on disease in experimental infection. ped virus was isolated from the small intestine of piglets inoculated with ped samples and cultured in vero, porcine bladder and kidney cells propagated in collagen-coated tissue culture plates in maintenance medium (mm) containing trypsin. in porcine bladder and kidney cell cultures inoculated with isolated ped virus, cytop ... | 2000 | 10727829 |
| detection of porcine epidemic diarrhea virus by immunohistochemistry with recombinant antibody produced in phages. | several diagnostic methods including immunofluorescence, enzyme-linked immunosorbent assay, polymerase chain reaction and immunohistochemistry have been developed for the detection of porcine epidemic diarrhea virus (pedv). an immunohistochemical method using a new recombinant antibody produced by a phage antibody system (pas16) kit was investigated and compared with that using a monoclonal antibody for pedv detection in pedv-infected piglets. in both the immunohistochemical methods, pedv antige ... | 2000 | 10770610 |
| detection and differentiation of porcine epidemic diarrhoea virus and transmissible gastroenteritis virus in clinical samples by multiplex rt-pcr. | a multiplex reverse-transcriptase-pcr (rt-pcr) procedure was developed for the simultaneous detection of porcine epidemic diarrhoea virus (pedv) and transmissible gastroenteritis virus (tgev) in preweaning pigs with diarrhoea. the membrane gene of pedv and the nucleocapsid gene of tgev were chosen as targets. the pcr products of pedv and tgev had molecular sizes of 412 and 612 base pairs, respectively. primers from pedv did not react with any tgev tested and vice versa. in addition, the primers ... | 2000 | 10872784 |
| immunoprophylactic effect of chicken egg yolk immunoglobulin (ig y) against porcine epidemic diarrhea virus (pedv) in piglets. | porcine epidemic diarrhea virus (pedv) is the causative agent of neonatal diarrhea in piglets, which causes high mortality rates. in this study, the immunoprophylactic effects of chicken egg yolk immunoglobulin (ig y) against pedv were investigated in neonatal pigs. ig y was found to reduce the mortality in piglets after challenge exposures. the field application of ig y also revealed significant differences in survival rates of piglets given ig y, as compared with placebo or control. the result ... | 2000 | 11039591 |
| prevalence of porcine epidemic diarrhoea virus and transmissible gastroenteritis virus infection in korean pigs. | | 2000 | 11110482 |
| dual infection with pcv-2 and porcine epidemic diarrhoea virus in neonatal piglets. | | 2001 | 11334076 |
| differential detection of transmissible gastroenteritis virus and porcine epidemic diarrhea virus by duplex rt-pcr. | transmissible gastroenteritis (tge) and porcine epidemic diarrhea (ped) are highly contagious enteric diseases of piglets. the clinical signs of these diseases are very similar and include watery, yellowish diarrhea. thus, the effective differential detection of tge virus and ped virus is required. in the present study, a duplex reverse transcription-polymerase chain reaction (rt-pcr) was established for the differential detection of tge and ped viruses. the primers were designed for the s gene ... | 2001 | 11724144 |
| a serodiagnostic elisa using recombinant antigen of swine transmissible gastroenteritis virus nucleoprotein. | a serodiagnostic elisa utilizing the recombinant nucleoprotein (rn protein) of transmissible gastroenteritis virus (tgev) was developed, and evaluated by examining a panel of 141 virus neutralization (vn) positive and 101 negative sera. the rn protein-based elisa (rnelisa) appeared to be highly sensitive and specific (98.6% and 98.0%, respectively) when it was compared to the vn test. the result was similar to that of an elisa based on purified viral antigens with showing good correlation (r=0.8 ... | 2001 | 11767065 |
| isotype-specific antibody-secreting cells in systemic and mucosal associated lymphoid tissues and antibody responses in serum of conventional pigs inoculated with pedv. | an enzyme-linked immunospot (elispot) has been developed to detect porcine epidemic diarrhea virus (pedv)-specific antibody secreting cells (asc) in gut associated lymphoid tissues (duodenum and ileum lamina propria and mesenteric lymph nodes) and systemic locations (spleen and blood) of conventional pigs so as to characterise the mucosal and systemic antibody response generated by the infection with pedv. a total number of 28 eleven-day-old conventional pigs were orally inoculated with the fiel ... | 2002 | 11825594 |
| mucosal and systemic isotype-specific antibody responses and protection in conventional pigs exposed to virulent or attenuated porcine epidemic diarrhoea virus. | eleven-day-old conventionally reared piglets were inoculated orally with two different doses of the cell-culture adapted strain cv-777 of the porcine epidemic diarrhoea virus (pedv) or the virulent isolate of the same strain and challenged with the same virulent pedv 3 weeks later. pigs inoculated with the two doses of the attenuated virus did not show any typical sign of the disease, and virus shedding was not frequent. in contrast, 31% of pigs exposed to the virulent pedv developed diarrhoea a ... | 2002 | 11867170 |
| comparison of reverse transcription polymerase chain reaction, immunohistochemistry, and in situ hybridization for the detection of porcine epidemic diarrhea virus in pigs. | reverse transcription polymerase chain reaction (rt-pcr), immunohistochemistry, and in situ hybridization were compared for the detection of porcine epidemic diarrhea virus (pedv). fifteen piglets experimentally infected with pedv were used in the study. in addition, 94 diarrheic piglets submitted to the department of veterinary pathology in seoul national university for diagnosis of pedv infection were used to compare the 3 methods. antigen and nucleic acid of pedv were detected in 15/15, 13/15 ... | 2002 | 11989732 |
| the propagation of a porcine epidemic diarrhea virus in swine cell lines. | a strain of porcine epidemic diarrhea virus (pedv), p-5v, utilized as a live virus vaccine in japan was infected to a swine cell lines, ksek6 and ib-rs-2 cells. clear cpe, characterized by cellular destruction, started to appear in the infected cells on 2-3 days post infection (dpi) and affected cells was completely degenerated on 4 dpi. the virus was serially passaged in the cells even without addition of trypsin. small but clear plaques were formed under an agar overlay medium on the cells. th ... | 2002 | 12173769 |
| lymphoproliferative responses and protection in conventional piglets inoculated orally with virulent or attenuated porcine epidemic diarrhoea virus. | lymphocyte proliferative responses were evaluated in mucosal (mesenteric lymph nodes) and systemic (spleen and blood) lymphoid tissues of conventional piglets inoculated with the virulent or attenuated isolates of porcine epidemic diarrhoea virus (pedv) strain cv-777 and challenged 21 days later with the virulent isolate of the same virus. a lymphoproliferative assay was developed in which mononuclear cells isolated from lymphoid tissues at different postinoculation and postchallenge days underw ... | 2002 | 12176140 |
| identification of the epitope region capable of inducing neutralizing antibodies against the porcine epidemic diarrhea virus. | in order to identify the neutralizing epitope of the porcine epidemic diarrhea virus (pedv), the spike protein region that is presumed to contain the virus-neutralizing epitope was determined. this was based on the sequence information for the neutralizing epitope of the transmissible gastroenteritis virus (tgev). a recombinant protein that corresponds to the spike region of tgev was produced, and polyclonal antisera were generated using the recombinant protein. it was discovered that polyclonal ... | 2002 | 12442904 |
| differentiation between porcine epidemic diarrhea virus and transmissible gastroenteritis virus in formalin-fixed paraffin-embedded tissues by multiplex rt-nested pcr and comparison with in situ hybridization. | porcine epidemic diarrhea virus (pedv) and transmissible gastroenteritis virus (tgev) were detected and differentiated in formalin-fixed, paraffin-embedded tissues from experimentally and naturally infected pigs by multiplex reverse transcription-nested polymerase chain reaction (rt-npcr). the results of this new method were compared with in situ hybridization. a method based on xylene deparaffinization followed by proteinase k digestion yielded rna of a suitable quality for reliable and consist ... | 2003 | 12565152 |
| differentiation of a vero cell adapted porcine epidemic diarrhea virus from korean field strains by restriction fragment length polymorphism analysis of orf 3. | a porcine epidemic diarrhea virus (pedv) designated dr13 was isolated in vero cells and serially passaged by level 100. the virus was titrated at regular intervals of the passage level. open reading frame (orf) 3 sequences of the virus at passage levels 20, 40, 60, 80, and 100 were aligned and compared using a computer software program. suitability of the restriction fragment length polymorphism (rflp) analysis for differentiating the virus from other korean field strains was investigated. the d ... | 2003 | 12706667 |
| cloning and sequence analysis of the nucleocapsid gene of porcine epidemic diarrhea virus chinju99. | the nucleocapsid (n) gene of the porcine epidemic diarrhea virus (pedv) chinju99 which was previously isolated in chinju, korea was cloned and sequenced to establish the information for the development of genetically engineered diagnostic reagents. also, sequences of the nucleotides and deduced amino acids of the chinju99 n gene were analyzed by alignment with those of cv777 and brl/87. the nucleotide sequence encoding the entire n gene open reading frame (orf) of chinju99 was 1326 bases long an ... | 2003 | 12803473 |
| experimental infection of piglets with a korean strain of porcine epidemic diarrhoea virus. | thirty colostrum-deprived piglets aged 1 day were inoculated with a korean strain of porcine epidemic diarrhoea virus (pedv). the purpose was to elucidate the pathogenicity and viral distribution in pedv-infected piglets over a period of 60 h, by morphometric analysis, in-situ hybridization and immunohistochemistry. at 24-60 h post-inoculation (hpi), the villous height/crypt depth (vh/cd) ratio of infected pigs was significantly less than that of control pigs. positive cells typically exhibited ... | 2003 | 12859908 |
| cloning and sequence analysis of the spike gene of porcine epidemic diarrhea virus chinju99. | the spike (s) gene of the porcine epidemic diarrhea virus (pedv) chinju99 which was previously isolated in chinju, korea was cloned and sequenced to aid in the development of genetically engineered vaccines and diagnostic reagents against pedv. the nucleotide sequence encoding the entire s gene open reading frame (orf) of chinju99 was 4,152 bases long encoding 1,383 amino acids. it consisted of 1,001 adenine (24.1%), 849 cytosine (20.4%), 877 guanine (21.1%) and 1425 thymine (34.3%) residues. th ... | 2003 | 12876452 |
| detection of porcine circovirus type 2 in feces of pigs with or without enteric disease by polymerase chain reaction. | to establish the sensitive polymerase chain reaction(pcr) method and detect porcine circovirus type 2 (pcv2) from intestines and feces of commercial swine herds with or without enteric disease, intestinal samples from 68 pigs and 29 fecal samples from commercial swine farms were collected. a primer set, forward primer 5'-gaagaatggaagaagcgg-3' and reverse primer 5'-ctcacagcagtagacaggt-3', could detect the virus at a concentration as low as 2 infectious virions per milliliter under controlled cond ... | 2003 | 12918820 |
| induction of antigen-specific systemic and mucosal immune responses by feeding animals transgenic plants expressing the antigen. | a report from that the presence of lactogenic immunity in pigs protected suckling piglets from porcine epidemic diarrhea virus (pedv) infection suggested that inducing mucosal immune responses in lactating pigs is an effective way of protecting swine from pedv infection. in this study, we developed transgenic tobacco plants that express the antigen protein corresponding to the neutralizing epitope of pedv spike protein, and tested whether feeding the plants to pigs induced an effective immune re ... | 2003 | 12922142 |
| 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 ... | 2003 | 12934944 |
| application of reverse transcription polymerase chain reaction to detect porcine epidemic diarrhea virus in vero cell culture. | | 1999 | 12968738 |
| monoclonal antibody-based immunohistochemical detection of porcine epidemic diarrhea virus antigen in formalin-fixed, paraffin-embedded intestinal tissues. | an immunohistochemistry technique was developed for the diagnosis of porcine epidemic diarrhea virus (pedv). the technique was tested on formalin-fixed, paraffin-embedded intestinal tissues from piglets naturally infected with pedv. five different monoclonal antibodies (mabs) were tested in this study. pedv antigen was consistently detected in the plp (4% paraformaldehyde, 100 mm l-lysine dihydrochloride, 10 mm sodium m-periodate in phosphate-buffered saline)-fixed pedv-infected vero cells or fo ... | 1999 | 12968761 |
| identification of a putative cellular receptor 150 kda polypeptide for porcine epidemic diarrhea virus in porcine enterocytes. | porcine epidemic diarrhea virus (pedv) causes an acute enteritis in pigs of all ages, often fatality for neonates. pedv occupies an intermediate position between two well characterized members of the coronavirus group i, human coronavirus (hcov-229e)and transmissible gastroenteritis virus (tgev) which uses aminopeptidase n (apn), a 150 kda protein, as their receptors. however, the receptor of the pedv has not been identified yet. a virus overlay protein binding assay (vopba) was used to identify ... | 2003 | 14685034 |
| effect of temperature on the detection of porcine epidemic diarrhea virus and transmissible gastroenteritis virus in fecal samples by reverse transcription-polymerase chain reaction. | the effect of storage temperature was determined for the detection of porcine epidemic diarrhea virus (pedv) and transmissible gastroenteritis virus (tgev) in fecal samples from experimentally and naturally infected pigs by multiplex reverse transcription-polymerase chain reaction (rt-pcr). to examine the effect of storage temperature on the ability to detect pedv and tgev rna by multiplex rt-pcr, fecal samples were stored for different temperatures (4, 21, 36, and 45 c) before extracting viral ... | 2004 | 15152841 |
| verification of sensitivity and specificity of group a rotavirus detection in piglets faeces with monoclonal blocking elisa methods. | monoclonal antibodies to group a rotavirus vp6 protein were prepared and used for verification of three blocking enzyme-linked immunosorbent assay (elisa) modifications to detect rotavirus a. selected competitive blocking elisa (cb-elisa) and electron microscopy (em) were used for examination of 194 field faecal samples of piglets affected with diarrhoea. rotavirus was detected in 43 samples (22.2%) by cb-elisa method, whereas in 26 (13.4%) samples by em examination. however, of 26 samples posit ... | 2004 | 15228549 |
| high-level expression of the neutralizing epitope of porcine epidemic diarrhea virus by a tobacco mosaic virus-based vector. | porcine epidemic diarrhea virus (pedv) causes acute enteritis in pigs of all ages and is often fatal for neonates. a tobacco mosaic virus (tmv)-based vector was utilized for the expression of a core neutralizing epitope of pedv (coe) for the development of a plant-based vaccine. in this study, the coding sequence of a coe gene was optimized based on the modification of codon usage in tobacco plant genes and the removal of mrna-destabilizing sequences. the native and synthetic coe genes were clon ... | 2004 | 15477091 |
| testing the hypothesis of a recombinant origin of the sars-associated coronavirus. | the origin of severe acute respiratory syndrome-associated corona-virus (sars-cov) is still a matter of speculation, although more than one year has passed since the onset of the sars outbreak. in this study, we implemented a 3-step strategy to test the intriguing hypothesis that sars-cov might have been derived from a recombinant virus. first, we blasted the whole sars-cov genome against a virus database to search viruses of interest. second, we employed 7 recombination detection techniques wel ... | 2005 | 15480857 |
| an elisa optimized for porcine epidemic diarrhoea virus detection in faeces. | monoclonal antibodies to porcine epidemic diarrhoea virus (pedv) membrane protein m were prepared and used for the comparative assessment of three blocking elisa variants to detect pedv. the competitive blocking elisa (cb-elisa) format showed the highest sensitivity, allowing detection of 10(2.5) plaque-forming units of pedv/ml in culture medium. its specificity was verified by inclusion of control samples containing transmissible gastroenteritis virus (tgev) and rotavirus a in each analysis. ei ... | 2004 | 15607079 |
| rt-pcr-based dot blot hybridization for the detection and differentiation between porcine epidemic diarrhea virus and transmissible gastroenteritis virus in fecal samples using a non-radioactive digoxigenin cdna probe. | multiplex reverse transcription-polymerase chain reaction (rt-pcr)-based dot blot hybridization was developed to increase the sensitivity for the detection and differentiation between porcine epidemic diarrhea virus (pedv) and transmissible gastroenteritis virus (tgev) in fecal samples. fecal samples found positive by rt-pcr-based agarose gel electrophoresis were always found positive by rt-pcr-based dot blot hybridization. in addition, 5 out of 10 fecal samples which were negative for pedv by r ... | 2005 | 15620395 |
| structure-based preliminary analysis of immunity and virulence of sars coronavirus. | the research on sars-associated coronavirus (sars-cov) has not stopped since its discovery, but the pathogenesis of sars is still unclear. to explore the possible molecular mechanisms of the invasion and virulence of sars-cov, we investigated the structural basis of the viral proteins using computational biology. forty-five motifs relating to superantigens, toxins and other bioactive molecules were detected in the proteins of sars-cov. the results showed that the distribution of the motifs varie ... | 2004 | 15671749 |
| cloning and sequence analysis of the m gene of porcine epidemic diarrhea virus ljb/03. | porcine epidemic diarrhea virus (pedv) ljb/03 was isolated from the fece of piglets infected with pedv on a pig farm, heilongjiang province, china. the m gene of ljb/03 was amplified from the rna extracted directly from the fece samples by rt-pcr and cloned into pmd18-t vector. the m gene cdna was sequenced and encompasses an open reading frame of 681 nucleotides, encoding a 226-amino acid protein. the ljb/03 m gene has a base composition of 152 adenines (22%), 153 cytosines (23%), 161 guanines ... | 2005 | 15744564 |
| expression of the synthetic neutralizing epitope gene of porcine epidemic diarrhea virus in tobacco plants without nicotine. | porcine epidemic diarrhea virus (pedv) causes acute diarrhea and dehydration in pigs, leading to death with a high mortality. in this study, the synthetic neutralizing epitope gene of pedv, which was optimized based on the coding sequence of tobacco plant genes being modified, was expressed in the no-nicotine tobacco plants. the synthetic gene was cloned into the plant expression vector under the control of camv 35s promoter and transformed by an agrobacterium-mediated transformation. the amount ... | 2005 | 15755614 |
| mixed infections in vitro with different chlamydiaceae strains and a cell culture adapted porcine epidemic diarrhea virus. | assuming a synergistic or additive effect of chlamydiaceae in coexistence with other enteropathogenic agents, the viral/bacterial interaction between a cell culture adapted porcine epidemic diarrhea virus (ca-pedv) and different chlamydiaceae strains was studied in vitro. vero cells were dually infected with ca-pedv and one of the three chlamydial strains chlamydia trachomatis s45, chlamydophila abortus s26/3 or chlamydophila pecorum 1710s. three experimental protocols were designed varying the ... | 2005 | 15778027 |
| cloning and sequence analysis of the korean strain of spike gene of porcine epidemic diarrhea virus and expression of its neutralizing epitope in plants. | porcine epidemic diarrhea virus (pedv) causes acute diarrhea and dehydration in pigs and leads to death with a high mortality rate, which has been reported notably in korea. the spike (s) gene of the pedv isolated in korea was cloned and sequenced. the nucleotide sequence encoding the entire s gene open reading frame of korean strain was 4161 bases long encoding 1387 amino acids. the neutralizing epitope of korean pedv (k-coe) was expressed in tobacco plants using agrobacterium-mediated protein ... | 2005 | 15866725 |
| expression of synthetic neutralizing epitope of porcine epidemic diarrhea virus fused with synthetic b subunit of escherichia coli heat-labile enterotoxin in tobacco plants. | the pentameric b subunit of escherichia coli heat-labile enterotoxin (ltb) can be used as an efficient mucosal carrier of either immunogenic or tolerogenic t-cell epitopes. co-delivery of therapeutic proteins with carrier proteins could increase the effectiveness of the antigen. this paper reports the ability of transgenic tobacco plants to express a fusion protein consisting of the synthetic ltb and a synthetic neutralizing epitope of porcine epidemic diarrhea virus (pedv), causing an enteric d ... | 2006 | 16174563 |
| comparison of an enzyme-linked immunosorbent assay with serum neutralization test for serodiagnosis of porcine epidemic diarrhea virus infection. | an indirect porcine epidemic diarrhea (ped) virus (pedv) enzyme-linked immunosorbent assay (elisa) was compared with the serum neutralization (sn) test by testing 46 samples from experimentally infected sows, 73 samples from naive sows, and 1,024 field sow samples from 48 commercial swine farms of undefined ped status. the sn test and the elisa were performed using pedv, kpedv-9 strain. viral proteins as a coating antigen of pedv elisa were extracted from the cytoplasm of pedv-infected vero cell ... | 2005 | 16294000 |
| use of an internal control in a quantitative rt-pcr assay for quantitation of porcine epidemic diarrhea virus shedding in pigs. | porcine epidemic diarrhea virus (pedv), a member of the family coronaviridae, has caused a devastating enteric disease in the korean swine industry. previously, the differences between virulent field pedv strains and a vero cell culture adapted pedv dr13 strain were determined using restriction fragment length polymorphism analysis (rflp), and pedv shedding patterns in pigs were reported. in an extension to these studies, an internal control was constructed and quantitative analysis of virus she ... | 2006 | 16300838 |
| identification of porcine circovirus type 2 in retrospective cases of pigs naturally infected with porcine epidemic diarrhoea virus. | the identification of porcine circovirus type 2 (pcv2) was studied in fresh intestinal tissues by polymerase chain reaction (pcr) and in formalin-fixed, paraffin-wax-embedded intestinal tissues by in situ hybridisation. the tissues came from pigs naturally infected with porcine epidemic diarrhoea virus (pedv). a total of 35 (32.7%) of 107 small intestinal samples from pigs naturally infected with pedv were found to be positive using pcr. positive signals for pcv2 were detected in 32 (29.9%) of 1 ... | 2006 | 16427593 |
| the effects of transplacental porcine circovirus type 2 infection on porcine epidemic diarrhoea virus-induced enteritis in preweaning piglets. | the effects of transplacental porcine circovirus type 2 (pcv2) infection on porcine epidemic diarrhoea virus (pedv)-induced enteritis were examined in neonatal piglets. six pregnant sows were randomly allocated to an infected (n=3) or control group (n=3). three pregnant sows were inoculated intranasally with 6 ml of tissue culture fluid containing 1.2 x 10(5) tissue culture infective doses 50% (tcid(50))/ml of pcv2 strain snuvr000470 three weeks before the expected farrowing date. three control ... | 2006 | 16624710 |
| oral efficacy of vero cell attenuated porcine epidemic diarrhea virus dr13 strain. | a vero cell attenuated porcine epidemic diarrhea virus (pedv) strain, dr13, was distinguished from wild-type pedv using restriction enzyme fragment length polymorphism (rflp). cell attenuated dr13 was orally or intramuscularly (im) administered to late-term pregnant sows, and mortality resulting from the highly virulent pedv challenge was investigated in passively immunized suckling piglets of the two different groups. the mortality rate of the oral group (13%) was lower than that of the im grou ... | 2007 | 16730762 |
| decreased activity of brush border membrane-bound digestive enzymes in small intestines from pigs experimentally infected with porcine epidemic diarrhea virus. | brush border membrane-bound digestive enzymes such as disaccharidases (lactase, sucrase, and maltase), leucine aminopeptidase n, and alkaline phosphatase were measured in jejunum from pigs experimentally infected with porcine epidemic diarrhea virus (pedv). three piglets from the infected and control groups were euthanized by electrocution and subjected to necropsy at 24, 36, 48, 60, and 72 hours post-inoculation (hpi). the infection of pedv to jejunum resulted in significant decreases in brush ... | 2006 | 16759679 |
| multiplex reverse transcription-pcr for rapid differential detection of porcine epidemic diarrhea virus, transmissible gastroenteritis virus, and porcine group a rotavirus. | a novel multiplex reverse transcription polymerase chain reaction (multiplex rt-pcr) that can detect porcine epidemic diarrhea virus (pedv), transmissible gastroenteritis virus (tgev), and porcine group a rotavirus (gar) was developed. the 3 viruses (pedv, tgev, and porcine gar) are major agents in viral enteric diseases of piglets. as the clinical signs of these diseases are similar, including watery diarrhea, differential detection is required for etiologic diagnosis. a mixture of 3 pairs of p ... | 2006 | 16789718 |
| cloning and sequence analysis of the n gene of porcine epidemic diarrhea virus ljb/03. | the nucleocapsid (n) gene of the porcine epidemic diarrhea virus (pedv) strain ljb/03 which was previously isolated in heilongjiang province, china, was cloned, sequenced and compared with published sequences of other avian and mammalian coronavirus. the nucleotide sequence encoding the entire n gene open reading frame (orf) of ljb/03 was 1326 bases long and encoded a protein of 441 amino acids with predicted mr of 49 kda. it consisted of 405 adenines (30.5%), 294 cytosines (22.1%), 329 guanines ... | 2006 | 16972037 |
| cloning and further sequence analysis of the spike gene of attenuated porcine epidemic diarrhea virus dr13. | the spike (s) gene of the attenuated porcine epidemic diarrhea virus (pedv) dr13 was cloned and sequenced to further explore the functions of wild type pedv and attenuated pedv. sequencing revealed a single large orf of 4,149 nucleotides encoding a protein of 1,382 amino acids with predicted m(r) of 151 kda. the coding region of the s gene of attenuated pedv dr13 had 20 nucleotide changes that appeared to be significant determinants of function in that they produced changes in its predicted amin ... | 2007 | 17072758 |
| human coronavirus 229e encodes a single orf4 protein between the spike and the envelope genes. | the genome of coronaviruses contains structural and non-structural genes, including several so-called accessory genes. all group 1b coronaviruses encode a single accessory protein between the spike and envelope genes, except for human coronavirus (hcov) 229e. the prototype virus has a split gene, encoding the putative orf4a and orf4b proteins. to determine whether primary hcov-229e isolates exhibit this unusual genome organization, we analyzed the orf4a/b region of five current clinical isolates ... | 2006 | 17194306 |
| sequence analysis of the partial spike glycoprotein gene of porcine epidemic diarrhea viruses isolated in korea. | porcine epidemic diarrhea virus (pedv) causes a devastating enteric disease with acute diarrhea, dehydration and significant mortality in swine, thereby incurring heavy economic losses in korea. spike (s) glycoprotein has been suggested as an important determinant for pedv biological properties. in this study, the nucleotide and deduced amino acid sequences of the partial s glycoprotein genes of korean pedv isolates, including epitope region that is capable of inducing pedv-neutralizing antibodi ... | 2007 | 17436070 |
| porcine aminopeptidase n is a functional receptor for the pedv coronavirus. | porcine epidemic diarrhea virus (pedv) causes lethal diarrhea in piglets that leads to great economic losses in east asia. it was reported that aminopeptidase n (apn) is the receptor for transmissible gastroenteritis virus (tgev), human coronavirus 229e (hcov-229e) and feline coronavirus (fecov) which all belong to group i coronavirus including as well as pedv. it was also confirmed previously that porcine aminopeptidase n (papn) can bind to pedv, and anti-papn antibodies may inhibit the combina ... | 2007 | 17467767 |
| high-level prokaryotic expression of envelope exterior of membrane protein of porcine epidemic diarrhea virus. | the truncated fragment m' gene, encoding the exterior of the viral envelope protein of pedv, was subcloned into prokaryotic expression vector pgex-6p-1. the recombinant plasmid pgex-6p-m' was constructed and transformed into e. coli bl21(de3)plyss for expression. sds-page analysis showed recombinant truncated m' protein was highly expressed by pgex-6p-m' and the product fusion protein gst-m' reached 45% in the total bacteria proteins with the analysis of software alphaimager2200. the preliminary ... | 2007 | 17475420 |
| effects of epidermal growth factor on atrophic enteritis in piglets induced by experimental porcine epidemic diarrhoea virus. | epidermal growth factor (egf) promotes gastrointestinal mucosal recovery by stimulating the mitogenic activity of intestinal crypt epithelial cells. the aim of this study was to determine the effects of egf on atrophic enteritis induced in piglets by experimental infection with porcine epidemic diarrhoea virus (pedv) strain dr13. two groups of 12 conventional, colostrum-deprived, 1-day-old, large white-duroc cross breed piglets were inoculated orally with pedv (3 x 10(5) 50% tissue culture infec ... | 2008 | 17574457 |
| application of a focus formation assay for detection and titration of porcine epidemic diarrhea virus. | a focus formation assay (ffa) for detection and titration of porcine epidemic diarrhea virus (pedv) in a micro-culture system using vero cells and pap staining technique was evaluated. a linear correlation between the virus dilution and virus titer determined by ffa was observed between the range of 10 and 30 foci per well. comparative analysis between ffa and plaque assay showed no significant difference in estimating the titer of cell adapted pedv. however, the culture time required for detect ... | 2007 | 17576016 |
| development of a simple and rapid immunochromatographic strip test for diarrhea-causative porcine rotavirus in swine stool. | a rapid and simple immunochromatography (ic) strip test, for specific detection of porcine rotavirus (prv) in stool specimen, was developed. monoclonal antibodies (mabs) to the osu strain of prv have been produced in mice. among them, two hybridoma clones that generate mab-1 and mab-2, respectively, specific for vp6 protein of prv, have been selected. in the ic configuration, mab-1, one of the selected mabs was used to the designed coat microparticles (mp), while another mab-2 was used to fix it ... | 2007 | 17644198 |
| multiplex real-time rt-pcr for the simultaneous detection and quantification of transmissible gastroenteritis virus and porcine epidemic diarrhea virus. | transmissible gastroenteritis virus (tgev) and porcine epidemic diarrhea virus (pedv) are major etiological agents of diarrhea and death in piglets. multiplex real-time reverse transcriptase (rt)-pcr was developed for simultaneous differential quantification of each virus in a single reaction tube, using cy5- and fam-labeled taqman-probes based on sequences from the tgev and pedv nucleocapsid genes. the copy numbers for transcripts of tgev and pedv were quantified using this assay over a range f ... | 2007 | 17697717 |
| impact of porcine group a rotavirus co-infection on porcine epidemic diarrhea virus pathogenicity in piglets. | porcine epidemic diarrhea virus (pedv) and porcine group a rotavirus (pgar) are the main causative agents of acute diarrhea in piglets. in south korea, pgar is prevalent in piglets naturally infected with pedv. piglets naturally co-infected with pedv and pgar appeared to have severe and prolonged diarrhea that was distinct from that commonly observed. the aim of this study was to determine the impact of pgar co-infection on pedv pathogenicity in piglets. thirty-six colostrum-deprived, one-day ol ... | 2008 | 17727905 |
| genetic differentiation of the nucleocapsid protein of korean isolates of porcine epidemic diarrhoea virus by rt-pcr based restriction fragment length polymorphism analysis. | a reverse transcriptase polymerase chain reaction (rt-pcr) based restriction fragment length polymorphism (rflp) analysis based on the nucleocapsid (n) gene was developed to differentiate between field isolates of porcine epidemic diarrhoea virus (pedv) and a vaccine strain, j-vac. thirteen field isolates of pedv from korea were distinguishable from the vaccine strain and the prototype pedv strain cv777 by rflp using tru9i. rflp patterns in 11 of 13 field pedv isolates were different from the va ... | 2008 | 17728163 |
| cloning and further sequence analysis of the orf3 gene of wild- and attenuated-type porcine epidemic diarrhea viruses. | the open reading frame (orf3) genes of the parent dr13, attenuated dr13, kped-9, p-5v, and 12 field samples were cloned and sequenced to further explore the functions of wild- and attenuated-type porcine epidemic diarrhea viruses (pedvs). sequencing revealed that wild-type pedvs orf3 genes had a single orf of 675 nucleotides encoding a protein of 224 amino acids with a predicted m (r) of 25.1-25.3 kda. attenuated-type pedvs orf3 genes had a single orf of 624 nucleotides encoding a protein of 207 ... | 2008 | 17932736 |
| in vitro inhibition of coronavirus replications by the traditionally used medicinal herbal extracts, cimicifuga rhizoma, meliae cortex, coptidis rhizoma, and phellodendron cortex. | a search for new anti-coronaviral drugs to treat coronaviral infections was motivated by an outbreak of severe acute respiratory syndrome (sars). | 2008 | 18036887 |
| the gprlqpy motif located at the carboxy-terminal of the spike protein induces antibodies that neutralize porcine epidemic diarrhea virus. | the spike protein of porcine epidemic diarrhea virus is the main surface glycoprotein involved in virus attachment and entry and therefore is the target of neutralizing antibodies. here, the immunogenicity of a novel antigenic domain found on the carboxy-terminal of the spike protein characterized by the peptide motif gprlqpy, was evaluated. a synthetic peptide whose linear sequence is identical to the 24a.a. carboxy-terminal portion of the spike protein (s-ct24) elicited a strong antibody respo ... | 2008 | 18067984 |
| molecular characterization and phylogenetic analysis of membrane protein genes of porcine epidemic diarrhea virus isolates in china. | six porcine epidemic diarrhea viruses (pedvs) were isolated from the fecal samples of piglets infected with pedv in 2006 in china. the membrane (m) protein genes of six pedv isolates were amplified by reverse transcriptase-polymerase chain reaction (rt-pcr), then cloned, sequenced, and compared with each other as well as those ten pedv reference strains. the m protein genes of six chinese pedv isolates consisted of 692 nucleotides containing a single open reading frame (orf) of 681 nucleotides, ... | 2008 | 18214664 |
| epidemic of diarrhoea caused by porcine epidemic diarrhoea virus in italy. | there was an epidemic of diarrhoea affecting pigs of all ages in italy between may 2005 and june 2006. in 63 herds the cause was confirmed as porcine epidemic diarrhoea virus by electron microscopy, immunoelectron microscopy, pcr and serology. watery diarrhoea without mucus and blood was usually associated with a reduction of feed consumption. in farrowing-to-weaning herds, diarrhoea affected the sows and suckling piglets, and the mortality in newborn piglets was up to 34 per cent. in growers an ... | 2008 | 18326842 |
| identification of two novel b cell epitopes on porcine epidemic diarrhea virus spike protein. | s1d (residues 636-789) is a neutralizing epitope region on the spike protein (s) of porcine epidemic diarrhea virus (pedv). to accurately identify epitopes on s1d, the s1-phage library containing the gene encoding the s1d region of pedv s protein was micropanned by six specific monoclonal antibodies (mcabs) against the s1d region. these micropanned epitope regions (mer) were focused on 696-779 amino acids of the s protein. to further map epitopes of the mer, seven overlapping mini-fragments cove ... | 2008 | 18400422 |
| rapid detection of porcine reproductive and respiratory syndrome virus by reverse transcription loop-mediated isothermal amplification assay. | a rapid detection assay based on reverse transcription loop-mediated isothermal amplification (rt-lamp) has been developed for detecting porcine reproductive and respiratory syndrome virus (prrsv). the rt-lamp assay utilized a set of six primers to amplify the open reading frame 6 (orf6) of the prrsv. the amplified products were analyzed by agarose gel electrophoresis or visualized by colorimetric method. the results demonstrated that the rt-lamp assay detected all 22 different prrsv isolates, h ... | 2009 | 18926852 |
| antiviral activity of quercetin 7-rhamnoside against porcine epidemic diarrhea virus. | porcine epidemic diarrhea virus (pedv) is the predominant cause of severe entero-pathogenic diarrhea in swine. the lack of effective therapeutical treatment underlines the importance of research for new antivirals. in this study, we identified q7r, which actively inhibited pedv replication with a 50% inhibitory concentration (ic(50)) of 0.014 microg/ml. the 50% cytotoxicity concentration (cc(50)) of q7r was over 100 microg/ml and the derived therapeutic index was 7142. several structural analogu ... | 2009 | 18992773 |