antigenic relationship of the feline infections peritonitis virus to coronaviruses of other species.utilizing the direct and indirect fluorescent antibody procedure, the antigenic relationship of the feline infectious peritonitis virus (fipv) to 7 other human and animal coronaviruses was studied. fipv was found to be closely related to transmissible gastroenteritis virus (tgev) of swine. transmissible gastroenteritis virus and fipv were in turn antigenically related to human coronavirus 229e (hcv-229e) and canine coronavirus (ccv). an interesting finding in the study was that the 8 coronavirus ...197881044
human aminopeptidase n is a receptor for human coronavirus 229e.human coronaviruses (hcv) in two serogroups represented by hcv-229e and hcv-oc43 are an important cause of upper respiratory tract infections. here we report that human aminopeptidase n, a cell-surface metalloprotease on intestinal, lung and kidney epithelial cells, is a receptor for human coronavirus strain hcv-229e, but not for hcv-oc43. a monoclonal antibody, rbs, blocked hcv-229e virus infection of human lung fibroblasts, immunoprecipitated aminopeptidase n and inhibited its enzymatic activi ...19921350662
antigenic homology among coronaviruses related to transmissible gastroenteritis virus.the antigenic homology of 26 coronavirus isolates, of which 22 were antigenically related to transmissible gastroenteritis virus (tgev), was determined with 42 monoclonal antibodies. type, group, and interspecies specific epitopes were defined. two group specific mabs distinguished the enteric tgev isolates from the respiratory variants. an antigenic subsite involved in neutralization was conserved in porcine, feline, and canine coronavirus. the classification of the human coronavirus 229e in a ...19901689525
in vitro antiviral activity and preliminary clinical trials of a new adamantane compound.a compound, 1'-methyl spiro (adamantane-2,3'-pyrrolidine) maleate, chemically related to the antiviral drug amantadine, was tested for activity in vitro against a number of human respiratory viruses. by a variety of techniques, it was shown to be active against a wide range of human and animal influenza a viruses. the effect was, however, variable and ranged from high activity against two 1957 asian strains to no observable activity against a 1971 strain. like amantadine, the drug did not inhibi ...19734364762
detection of coronavirus 229e antibody by indirect hemagglutination.tannic-acid treated sheep erythrocytes (fresh or glutaraldehyde preserved) were sensitized with 229e antigens from human embryonic lung (ru-1) cell cultures. indirect hemagglutination (iha) antigen titers in 229e-infected cell cultures paralleled virus infectivity and complement fixation (cf) antigen titers. the identity of the iha antigen was confirmed by testing extracts from inoculated and control cell cultures for ability to inhibit iha. also, significant increases in iha antibody were demon ...19724674373
detection of human coronavirus 229e-specific antibodies using recombinant fusion proteins.human coronaviruses are known to be a common cause of respiratory infections in man. however, the diagnosis of human coronavirus infections is not carried out routinely, primarily because the isolation and propagation of these viruses in tissue culture is difficult and time consuming. the aim of this study was to evaluate the use of recombinant, bacterial expressed proteins in the serodiagnosis of coronavirus infections. two proteins were examined: the human coronavirus 229e nucleocapsid protein ...19958537456
feline aminopeptidase n is a receptor for all group i coronaviruses.human coronavirus hcv-229e and porcine transmissible gastroenteritis virus (tgev), both members of coronavirus group i, use aminopeptidase n (apn) as their cellular receptors. these viruses show marked species specificity in receptor utilization as they can only use apn of their respective species to initiate virus infection. feline and canine coronaviruses are also group i coronaviruses. to determine whether feline apn could serve as a receptor for feline coronaviruses (fcovs), we cloned the cd ...19989782266
adaptation of human enteric coronavirus to growth in cell lines.the existence of human enteric coronavirus (hec) has been debated since its first description in stool by electron microscopy (em) in 1975. needed to resolve the issue is its cultivation in readily available cell lines.199910073413
ribonucleoprotein of avian infectious bronchitis virus.the ribonucleoprotein (rnp) of avian infectious bronchitis virus (ibv) was examined by electron microscopy after shadowing with carbon/platinum. linear rnp strands up to 6.7 microns in length, from three ivb strains, were sensitive to both pancreatic rnase and to proteases. these strands were obtained from spontaneously disrupted complete particles but not from disrupted incomplete particles that lacked rnp. they were also released from nonidet p40-disrupted particles and could be isolated on su ...19816268741
purification and biophysical properties of human coronavirus 229e. 1976824815
persistent infection of human oligodendrocytic and neuroglial cell lines by human coronavirus 229e.human coronaviruses (hucv) cause common colds. previous reports suggest that these infectious agents may be neurotropic in humans, as they are for some mammals. with the long-term aim of providing experimental evidence for the neurotropism of hucv and the establishment of persistent infections in the nervous system, we have evaluated the susceptibility of various human neural cell lines to acute and persistent infection by hucv-229e. viral antigen, infectious virus progeny and viral rna were mon ...199910074187
a strategy for the generation of infectious rnas and autonomously replicating rnas based on the hcv 229e genome.a strategy to generate in vitro transcripts representing infectious rnas and autonomously replicating rnas based on the hcv 229e genome is presented. pcr-dnas were ligated in vitro, resulting in 27 kbp and 22 kbp ligation products. these dnas can now be transcribed in vitro and the rnas tested for infectivity and their ability to replicate.19989782291
molecular determinants of species specificity in the coronavirus receptor aminopeptidase n (cd13): influence of n-linked glycosylation.aminopeptidase n (apn), a 150-kda metalloprotease also called cd13, serves as a receptor for serologically related coronaviruses of humans (human coronavirus 229e [hcov-229e]), pigs, and cats. these virus-receptor interactions can be highly species specific; for example, the human coronavirus can use human apn (hapn) but not porcine apn (papn) as its cellular receptor, and porcine coronaviruses can use papn but not hapn. substitution of papn amino acids 283 to 290 into hapn for the corresponding ...200111559807
completion of the porcine epidemic diarrhoea coronavirus (pedv) genome sequence.the sequence of the replicase gene of porcine epidemic diarrhoea virus (pedv) has been determined. this completes the sequence of the entire genome of strain cv777, which was found to be 28,033 nucleotides (nt) in length (excluding the poly a-tail). a cloning strategy, which involves primers based on conserved regions in the predicted orf1 products from other coronaviruses whose genome sequence has been determined, was used to amplify the equivalent, but as yet unknown, sequence of pedv. primary ...200111724265
characterization of a papain-like cysteine-proteinase encoded by gene 1 of the human coronavirus hcv 229e.expression of the coronaviral gene 1 polyproteins, pp 1a and pp 1ab, involves a series of proteolytic events that are mediated by virus-encoded proteinases similar to cellular papain-like cysteine-proteinases and the 3c-like proteinases of picornaviruses. in this study, we have characterized, in vitro, the human coronavirus hcv 229e papain-like cysteine-proteinase pcp 1. we show that pcp 1 is able to mediate cleavage of an aminoterminal polypeptide, p9, from in vitro translation products represe ...19989782276
replication and transcription of hcv 229e replicons. p6.replicons based upon the human coronavirus 229e (hcv 229e) genome were transfected into hcv 229e infected cells. we demonstrate that a synthetic rna comprised of 646 nucloetides from the 5' end and 1465 nucloetides from the 3' end of the hcv 229e genome is replication competent. we conclude that the cis-acting elements necessary for replication are located in these 5' and 3' genomic regions. furthermore, we inserted the intergenic region of the hcv 229e nucleocapsid protein gene into this basic ...19989782271
involvement of aminopeptidase n (cd13) in infection of human neural cells by human coronavirus 229e.attachment to a cell surface receptor can be a major determinant of virus tropism. previous studies have shown that human respiratory coronavirus hcv-229e uses human aminopeptidase n (hapn [cd13]) as its cellular receptor for infection of lung fibroblasts. although human coronaviruses are recognized respiratory pathogens, occasional reports have suggested their possible neurotropism. we have previously shown that human neural cells, including glial cells in primary cultures, are susceptible to h ...19989658094
coronavirus 229e-related pneumonia in immunocompromised patients.coronaviruses strains 229e and oc43 have been associated with various respiratory illnesses ranging from the self-resolving common cold to severe pneumonia. although chronic underlying conditions are major determinants of severe respiratory virus infections, few data about coronavirus-related pneumonia in immunocompromised patients are available. here we report 2 well-documented cases of pneumonia related to coronavirus 229e, each with a different clinical presentation. diagnosis was made on the ...200313130404
relationship of sars-cov to other pathogenic rna viruses explored by tetranucleotide usage profiling.the exact origin of the cause of the severe acute respiratory syndrome (sars) is still an open question. the genomic sequence relationship of sars-cov with 30 different single-stranded rna (ssrna) viruses of various families was studied using two non-standard approaches. both approaches began with the vectorial profiling of the tetra-nucleotide usage pattern v for each virus. in approach one, a distance measure of a vector v, based on correlation coefficient was devised to construct a relationsh ...200314499005
characterization of determinants involved in the feline infectious peritonitis virus receptor function of feline aminopeptidase n.feline aminopeptidase n (fapn) is a major cell surface receptor for feline infectious peritonitis virus (fipv), transmissible gastroenteritis virus (tgev), human coronavirus 229e (hcv 229e) and canine coronavirus (ccv). by using chimeric molecules assembled from porcine, human and feline apn we have analysed the determinants involved in the coronavirus receptor function of fapn. our results show that amino acids 670-840 of fapn are critically involved in its fipv and tgev receptor function where ...19989634079
effective amplification of 20-kb dna by reverse transcription pcr.polymerase chain reaction has been applied to the amplification of long dna fragments from a variety of sources, including genomic, mitochondrial, and viral dnas. however, polymerase chain reaction amplification from cdna templates produced by reverse transcription has generally been restricted to products of less than 10 kilobases. in this paper, we report a system to effectively amplify fragments up to 20 kilobases from human coronavirus 229e genomic rna. we demonstrate that the integrity of t ...19979324942
infection of primary cultures of human neural cells by human coronaviruses 229e and oc43.we evaluated the ability of human coronaviruses to infect primary cultures of human neural cells. double immunofluorescence with antibodies to virus and cell markers showed infection of fetal astrocytes and of adult microglia and astrocytes by strain oc43. rna amplification revealed infection of fetal astrocytes, adult microglia, and a mixed culture of adult oligodendrocytes and astrocytes by strain 229e. infectious virus was released only from fetal astrocytes, with higher titers for oc43. huma ...19978985420
an 'elaborated' pseudoknot is required for high frequency frameshifting during translation of hcv 229e polymerase mrna.the rna polymerase gene (gene 1) of the human coronavirus 229e is approximately 20 kb in length and is located at the 5' end of the positive-strand genomic rna. the coding sequence of gene 1 is divided into two large open reading frames, orf1a and orf1b, that overlap by 43 nucleotides. in the region of the orf1a/orf1b overlap, the genomic rna displays two elements that are known to mediate (-1) ribosomal frameshifting. these are the slippery sequence, uuuaaac, and a 3' pseudoknot structure. by i ...19938290341
molecular analysis of the human coronavirus (strain 229e) genome.the nucleotide sequence of the human coronavirus strain 229e (hcv 229e) has been determined. this article describes the organization of the virus genome, the predicted viral gene products and the mechanisms which regulate viral gene expression. this information provides a basis to investigate the biology and pathogenesis of hcv.19938219814
neurotropism of human coronavirus 229e.the 299e prototype strain of human coronavirus (hcv-229e) has so far been mainly associated with infections of the respiratory tract. in the present study, we show evidence for infection of the central nervous system (cns) by hcv-229e, both in vitro and in vivo. various human cell lines of cns origin were tested for their susceptibility to infection by hcv-229e. production of viral antigens was monitored by indirect immunofluorescence with monoclonal antibodies and infectious progeny virions by ...19938209751
evaluation of nested polymerase chain methods for the detection of human coronaviruses 229e and oc43.currently, the diagnosis of human respiratory coronavirus infection is either slow or insensitive. this paper describes nested polymerase chain reaction assays for the detection of human coronaviruses oc43 and 229e. the specificity and sensitivity of the assays have been determined and they have been applied to the detection of the viruses in nasal aspirates. these assays are more rapid and sensitive than cell culture and may replace the latter as the diagnostic method of choice.19947877631
coronavirus isolates sk and sd from multiple sclerosis patients are serologically related to murine coronaviruses a59 and jhm and human coronavirus oc43, but not to human coronavirus 229e.two coronaviruses (sk and sd), isolated from fresh autopsy brain tissue from two multiple sclerosis patients, were compared with known human and murine coronaviruses. in plaque neutralization assays, antisera prepared against multiple sclerosis isolates sk and sd demonstrated significant cross-reactivity to each other and to murine coronavirus a59, weak cross-reactivity to murine coronavirus jhm, but no cross-reactivity to the human coronavirus 229e. antiserum to sk or sd failed to inhibit hemag ...19817241654
the human coronavirus 229e superfamily 1 helicase has rna and dna duplex-unwinding activities with 5'-to-3' polarity.the human coronavirus 229e replicase gene encodes a protein, p66hel, that contains a putative zinc finger structure linked to a putative superfamily (sf) 1 helicase. a histidine-tagged form of this protein, hel, was expressed using baculovirus vectors in insect cells. the purified recombinant protein had in vitro atpase activity that was strongly stimulated by poly(u), poly(dt), poly(c), and poly(da), but not by poly(g). the recombinant protein also had both rna and dna duplex-unwinding activiti ...200010917600
establishment and maintenance of a persistent infection of l132 cells by human coronavirus strain 229e.a persistent infection by human coronavirus 229e (hcv/229e) was established in a human continuous cell line (l132). following the initial infection with stock hcv/229e, several cultures were established of which two (hv1 and hv4) have been maintained by continuous passage for two years. these cultures have shed high titres of infectious virus continuously into the supernatant fluid since their initiation. the persistently infected cells were resistant to homologous super-infection but supported ...19816171237
survival characteristics of airborne human coronavirus 229e.the survival of airborne human coronavirus 229e (hcv/229e) was studied under different conditions of temperature (20 +/- 1 degree c and 6 +/- 1 degree c) and low (30 +/- 5%), medium (50 +/- 5%) or high (80 +/- 5%) relative humidities (rh). at 20 +/- 1 degree c, aerosolized hcv/229e was found to survive best at 50% rh with a half-life of 67.33 +/- 8.24 h while at 30% rh the virus half-life was 26.76 +/- 6.21 h. at 50% rh nearly 20% infectious virus was still detectable at 6 days. high rh at 20 +/ ...19852999318
sequence analysis of the nucleocapsid protein gene of human coronavirus 229e.human coronaviruses are important human pathogens and have also been implicated in multiple sclerosis. to further understand the molecular biology of human coronavirus 229e (hcv-229e), molecular cloning and sequence analysis of the viral rna have been initiated. following established protocols, the 3'-terminal 1732 nucleotides of the genome were sequenced. a large open reading frame encodes a 389 amino acid protein of 43,366 da, which is presumably the nucleocapsid protein. the predicted protein ...19892922924
cerebrospinal fluid antibodies to coronavirus in patients with parkinson's disease.the etiology of parkinson's disease remains unknown, and a search for environmental agents continues. in 1985, fishman induced infection of the basal ganglia by a coronavirus in mice. although coronavirus is recognized primarily as a respiratory pathogen in humans, its affinity for the basal ganglia led us to investigate its possible role in human parkinson's disease. the cerebrospinal fluid of normal controls (ctl) (n = 18), and patients with parkinson's disease (pd (n = 20) and other neurologi ...19921316552
viral replicase gene products suffice for coronavirus discontinuous transcription.we have used vaccinia virus as a vector to clone a 22.5-kbp cdna that represents the 5' and 3' ends of the human coronavirus 229e (hcov 229e) genome, the hcov 229e replicase gene, and a single reporter gene (coding for green fluorescent protein [gfp]) located downstream of a regulatory element for coronavirus mrna transcription. when rna transcribed from this cdna was transfected into bhk-21 cells, a small percentage of cells displayed strong fluorescence. a region of the mrna encoding gfp was a ...200111413334
proteolytic processing at the amino terminus of human coronavirus 229e gene 1-encoded polyproteins: identification of a papain-like proteinase and its substrate.expression of the coronavirus gene 1-encoded polyproteins, pp1a and pp1ab, is linked to a series of proteolytic events involving virus-encoded proteinases. in this study, we used transfection and immunoprecipitation assays to show that the human coronavirus 229e-encoded papain-like cysteine proteinase, pcp1, is responsible for the release of an amino-terminal protein, p9, from the gene 1-encoded polyproteins. the same protein, p9, has also been identified in virus-infected cells. furthermore, us ...19989444982
human coronavirus polyadenylated rna sequences in cerebrospinal fluid from multiple sclerosis patients.human coronaviruses, represented by the two prototype strains hcv-oc43 and hcv-229e, are important human respiratory pathogens, also associated with necrotizing enterocolitis. two previous studies, one describing the electron microscopic observation of doughnut-shaped particles, resembling coronaviruses, in a perivascular inflammatory lesion of brain tissue taken at autopsy from a multiple sclerosis patient, and the other one reporting the isolation of coronaviruses from the brains of two multip ...19979208420
characterization of functional domains in the human coronavirus hcv 229e receptor.human aminopeptidase n (hapn or cd13) and porcine aminopeptidase n (papn) are functional receptors for human coronavirus (hcv) 229e and porcine transmissible gastroenteritis virus (tgev), respectively. however, hapn cannot function as a receptor for tgev and papn cannot function as a receptor for hcv 229e. in this study, we constructed a series of chimeric hapn/papn genes and expressed the corresponding proteins in transfected cells. subsequently, we identified the chimeric proteins that can fun ...19968887485
major genetic marker of nidoviruses encodes a replicative endoribonuclease.coronaviruses are important pathogens that cause acute respiratory diseases in humans. replication of the approximately 30-kb positive-strand rna genome of coronaviruses and discontinuous synthesis of an extensive set of subgenome-length rnas (transcription) are mediated by the replicase-transcriptase, a barely characterized protein complex that comprises several cellular proteins and up to 16 viral subunits. the coronavirus replicase-transcriptase was recently predicted to contain rna-processin ...200415304651
human coronavirus 229e infects polarized airway epithelia from the apical surface.gene transfer to differentiated airway epithelia with existing viral vectors is very inefficient when they are applied to the apical surface. this largely reflects the polarized distribution of receptors on the basolateral surface. to identify new receptor-ligand interactions that might be used to redirect vectors to the apical surface, we investigated the process of infection of airway epithelial cells by human coronavirus 229e (hcov-229e), a common cause of respiratory tract infections. using ...200010982370
human coronavirus hcov-229e enters susceptible cells via the endocytic pathway. 200111774468
coronavirus main proteinase (3clpro) structure: basis for design of anti-sars drugs.a novel coronavirus has been identified as the causative agent of severe acute respiratory syndrome (sars). the viral main proteinase (mpro, also called 3clpro), which controls the activities of the coronavirus replication complex, is an attractive target for therapy. we determined crystal structures for human coronavirus (strain 229e) mpro and for an inhibitor complex of porcine coronavirus [transmissible gastroenteritis virus (tgev)] mpro, and we constructed a homology model for sars coronavir ...200312746549
detection of human coronavirus 229e in nasal specimens in large scale studies using an rt-pcr hybridization assay.a novel human coronavirus (hcov) was this year recognized as the etiological agent of the severe acute respiratory syndrome. two other hcov (hcov-229e and hcov-oc43) have been known for 30 years. hcov-229e has been recently involved in nosocomial respiratory viral infections in high-risk children. however, their diagnosis is not routinely performed. currently, reliable immunofluorescence and cell culture methodologies are not available. as part of a four-year epidemiological study in a pediatric ...200415051115
characterization of the human coronavirus 229e (hcv 229e) gene 1.the sequence of the hcv 229e gene 1 has been determined and compared with the homologous sequences of the murine hepatitis virus and the avian infectious bronchitis virus. the coding sequence of gene 1 is 20,273 nucleotides in length. within this coding region are two large open reading frames, orf 1a (4,086 codons) and orf 1b (2,687 codons) which overlap by 40 nucleotides. in the overlapping region, the genomic rna can be folded into a pseudoknot structure, an element which is known to mediate ...19938209774
genome organization of porcine epidemic diarrhoea 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 ...19938209771
characterization of the expression and immunogenicity of the ns4b protein of human coronavirus 229e.sequencing of complementary dnas prepared from various coronaviruses has revealed open reading frames encoding putative proteins that are yet to be characterized and are so far only described as nonstructural (ns). as a first step in the elucidation of its function, we characterized the expression and immunogenicity of the ns4b gene product from strain 229e of human coronavirus (hcv-229e), a respiratory virus with a neurotropic potential. the gene was cloned and expressed in bacteria. a fusion p ...19989933919
mutational analysis of aminopeptidase n, a receptor for several group 1 coronaviruses, identifies key determinants of viral host range.feline coronavirus (fcov), porcine transmissible gastroenteritis coronavirus (tgev), canine coronavirus (ccov), and human coronavirus hcov-229e, which belong to the group 1 coronavirus, use aminopeptidase n (apn) of their natural host and feline apn (fapn) as receptors. using mouse-feline apn chimeras, we identified three small, discontinuous regions, amino acids (aa) 288 to 290, aa 732 to 746 (called r1), and aa 764 to 788 (called r2) in fapn that determined the host ranges of these coronavirus ...200717093189
the effects of coronavirus on human nasal ciliated respiratory epithelium.human coronavirus (hcov) accounts for 15-30% of common colds, but only one case report has described the effect of a coronavirus infection, that was asymptomatic, on human respiratory epithelium. the authors examined the effects of infection with hcov on ciliary structure and function in healthy volunteers infected by intranasal inoculation with hcov 229e. a further four volunteers were sham infected with ultraviolet-inactivated virus. immediately before inoculation (day 0) and 3 days later (day ...200111829103
simultaneous detection of fourteen respiratory viruses in clinical specimens by two multiplex reverse transcription nested-pcr assays.there is a need for rapid, sensitive, and accurate diagnosis of lower respiratory tract infections in children, elderly, and immunocompromised patients, who are susceptible to serious complications. the multiplex rt-nested pcr assay has been used widely for simultaneous detection of non-related viruses involved in infectious diseases because of its high specificity and sensitivity. a new multiplex rt-pcr assay is described in this report. this approach includes nested primer sets targeted to con ...200414748074
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 ...200617194306
feline aminopeptidase n is not a functional receptor for avian infectious bronchitis virus.coronaviruses are an important cause of infectious diseases in humans, including severe acute respiratory syndrome (sars), and have the continued potential for emergence from animal species. a major factor in the host range of a coronavirus is its receptor utilization on host cells. in many cases, coronavirus-receptor interactions are well understood. however, a notable exception is the receptor utilization by group 3 coronaviruses, including avian infectious bronchitis virus (ibv). feline amino ...200717324273
coronaviruses in children, greece. 200717582904
t-cell epitopes in severe acute respiratory syndrome (sars) coronavirus spike protein elicit a specific t-cell immune response in patients who recover from sars.the immunogenicity of hla-a2-restricted t-cell epitopes in the s protein of the severe acute respiratory syndrome coronavirus (sars-cov) and of human coronavirus strain 229e (hcov-229e) was analyzed for the elicitation of a t-cell immune response in donors who had fully recovered from sars-cov infection. we employed online database analysis to compare the differences in the amino acid sequences of the homologous t epitopes of hcov-229e and sars-cov. the identified t-cell epitope peptides were sy ...200415140958
seroepidemiology of group i human coronaviruses in children.recently, several new human coronaviruses have been identified.200717889596
reverse genetics of coronaviruses using vaccinia virus this article, we describe the reverse genetic system that is based on the use of vaccinia virus cloning vectors. this system represents a generic approach to coronavirus reverse genetics and was first described for the generation of recombinant human coronavirus 229e representing a group i coronavirus. subsequently, the same approach has been used to generate recombinant avian infectious bronchitis coronavirus and, recently, recombinant mouse hepatitis virus, representing group iii and group ...200515609513
development of three multiplex rt-pcr assays for the detection of 12 respiratory rna viruses.three multiplex hemi-nested rt-pcr assays were developed to detect simultaneously 12 rna respiratory viruses: influenza viruses a, b and c, human respiratory syncytial virus (hrsv), human metapneumovirus (hmpv), parainfluenza virus types 1-4 (piv-1, -2, -3 and -4), human coronavirus oc43 and 229e (hcov) and rhinovirus (hrv). an internal amplification control was included in one of the rt-pcr assays. the rt-pcr multiplex 1 and the hemi-nested multiplex 1 detected 1 and 0.1 tcid50 of rsv a, respec ...200515847919
history and recent advances in coronavirus discovery.human coronaviruses, first characterized in the 1960s, are responsible for a substantial proportion of upper respiratory tract infections in children. since 2003, at least 5 new human coronaviruses have been identified, including the severe acute respiratory syndrome coronavirus, which caused significant morbidity and mortality. nl63, representing a group of newly identified group i coronaviruses that includes nl and the new haven coronavirus, has been identified worldwide. these viruses are ass ...200516378050
human coronavirus infection among children with acute lower respiratory tract infection in thailand.this study was performed to further identify the previously uncharacterized human coronavirus 229e (hcov-229e) and human coronavirus oc43 (hcov-oc43) in thailand by using the rt-pcr technique. in addition, we performed this study in order to delineate the prevalence, the potential clinical impacts and evaluation of the genetic characterization of this pathogen in young children who presented with acute lower respiratory tract infections (alri).200717139182
synthesis, crystal structure, structure-activity relationships, and antiviral activity of a potent sars coronavirus 3cl protease inhibitor.a potent sars coronavirus (cov) 3cl protease inhibitor (tg-0205221, ki = 53 nm) has been developed. tg-0205221 showed remarkable activity against sars cov and human coronavirus (hcov) 229e replications by reducing the viral titer by 4.7 log (at 5 microm) for sars cov and 5.2 log (at 1.25 microm) for hcov 229e. the crystal structure of tg-0205221 (resolution = 1.93 a) has revealed a unique binding mode comprising a covalent bond, hydrogen bonds, and numerous hydrophobic interactions. structural c ...200616884309
towards a coronavirus-based hiv multigene vaccine.human immunodeficiency virus (hiv) infection represents one of the major health threats in the developing world. the costly treatment of infected individuals with multiple highly efficient anti-hiv drugs is only affordable in industrialized countries. thus, an efficient vaccination strategy is required to prevent the further spread of the infection. the molecular biology of coronaviruses and particular features of the human coronavirus 229e (hcov 229e) indicate that hcov 229e-based vaccine vecto ...200617162377
human coronavirus nl63, a new respiratory virus.from the mid-1960s onwards, it was believed that only two human coronavirus species infect humans: hcov-229e and hcov-oc43. then, in 2003, a novel member of the coronavirus family was introduced into the human population: sars-cov, causing an aggressive lung disease. fortunately, this virus was soon expelled from the human population, but it quickly became clear that the human coronavirus group contains more members then previously assumed, with hcov-nl63 identified in 2004. despite its recent d ...200616911043
use of antibody avidity assays for diagnosis of severe acute respiratory syndrome coronavirus indirect immunofluorescent assay (euroimmun ag, luebeck, germany) was used to investigate the avidity of immunoglobulin g (igg), igm, iga, and total ig (iggam) antibody responses to severe acute respiratory syndrome coronavirus (sars cov) infections. serial serum samples from eight patients collected during the first, third, and ninth months after the onset of infection were evaluated. it was found that low-avidity igg antibodies were detected in 15/15 (100%), 1/5 (20%), and 0/8 (0%) serum sa ...200717881505
hcov-oc43-induced encephalitis is in part immune-mediated. 200617037592
the widening scope of the past 2 years, at least three distinct human coronaviruses have been discovered, including the etiological agent associated with severe acquired respiratory syndrome (sars). these recently discovered viruses, with the exception of the sars associated coronavirus (sars-cov), are likely to be common respiratory viruses and may be responsible for a substantial proportion of respiratory tract disease.200616470161
[antigenicity analysis of nucleocapsid proteins of 3 human coronaviruses sars-cov, 229e and oc43 with their monoclonal antibodies].to prepare and characterize monoclonal antibodies (mabs) against the recombinant nucleocapsid (n) protein of 3 human coronaviruses sars-cov, 229e and oc43 and study the antigenic relationship between the 3 n proteins.200616546729
human coronavirus 229e binds to cd13 in rafts and enters the cell through caveolae.cd13, a receptor for human coronavirus 229e (hcov-229e), was identified as a major component of the triton x-100-resistant membrane microdomain in human fibroblasts. the incubation of living fibroblasts with an anti-cd13 antibody on ice gave punctate labeling that was evenly distributed on the cell surface, but raising the temperature to 37 degrees c before fixation caused aggregation of the labeling. the aggregated labeling of cd13 colocalized with caveolin-1 in most cells. the hcov-229e virus ...200415280478
enhanced identification of viral and atypical bacterial pathogens in lower respiratory tract samples with nucleic acid amplification tests.the advantages of nucleic acid amplification tests (nat) over conventional methods for the detection of pathogens in lower respiratory tract samples have not been established. nat for respiratory pathogens were performed on 439 endotracheal tube (ett) and bronchoalveolar lavage (bal) samples. a potential pathogen was detected in 87 samples. of 22 samples that tested positive by conventional methods, 15 tested positive for the same pathogen by nat, 1 tested positive for a different pathogen, 2 ha ...200616555283
analysis of human coronavirus 229e spike and nucleoprotein genes demonstrates genetic drift between chronologically distinct strains.historically, coronaviruses have been recognized as a cause of minor respiratory infections in humans. however, the recent identification of three novel human coronaviruses, one causing severe acute respiratory syndrome (sars), has prompted further examination of these viruses. previous studies of geographically and chronologically distinct human coronavirus 229e (hcov-229e) isolates have found only limited variation within s gene nucleotide sequences. in contrast, analysis of the s genes of con ...200616603522
false-positive results in a recombinant severe acute respiratory syndrome-associated coronavirus (sars-cov) nucleocapsid enzyme-linked immunosorbent assay due to hcov-oc43 and hcov-229e rectified by western blotting with recombinant sars-cov spike polypeptide.using paired serum samples obtained from patients with illness associated with increases in anti-human coronavirus oc43 (hcov-oc43) or anti-hcov-229e antibodies, we examined the possibility of false-positive results detected in a recombinant severe acute respiratory syndrome (sars)-associated coronavirus (sars-cov) nucleocapsid protein immunoglobulin g enzyme-linked immunosorbent assay (elisa). three of the 21 and 1 of the 7 convalescent-phase serum samples from persons with increases in antibod ...200415583332
structure and dynamics of sars coronavirus main proteinase (mpro). 200617037602
mosaic structure of human coronavirus nl63, one thousand years of evolution.before the sars outbreak only two human coronaviruses (hcov) were known: hcov-oc43 and hcov-229e. with the discovery of sars-cov in 2003, a third family member was identified. soon thereafter, we described the fourth human coronavirus (hcov-nl63), a virus that has spread worldwide and is associated with croup in children. we report here the complete genome sequence of two hcov-nl63 clinical isolates, designated amsterdam 57 and amsterdam 496. the genomes are 27,538 and 27,550 nucleotides long, r ...200617054987
cells of human aminopeptidase n (cd13) transgenic mice are infected by human coronavirus-229e in vitro, but not in vivo.aminopeptidase n, or cd13, is a receptor for serologically related coronaviruses of humans, pigs, and cats. a mouse line transgenic for the receptor of human coronavirus-229e (hcov-229e) was created using human apn (hapn) cdna driven by a hapn promoter. hapn-transgenic mice expressed hapn mrna in the kidney, small intestine, liver, and lung. hapn protein was specifically expressed on epithelial cells of the proximal convoluted renal tubules, bronchi, alveolar sacs, and intestinal villi. the hapn ...200515840518
comparative host gene transcription by microarray analysis early after infection of the huh7 cell line by severe acute respiratory syndrome coronavirus and human coronavirus 229e.the pathogenesis of severe acute respiratory syndrome-associated coronavirus (sars-cov) at the cellular level is unclear. no human cell line was previously known to be susceptible to both sars-cov and other human coronaviruses. huh7 cells were found to be susceptible to both sars-cov, associated with sars, and human coronavirus 229e (hcov-229e), usually associated with the common cold. highly lytic and productive rates of infections within 48 h of inoculation were reproducible with both viruses. ...200515858003
cinanserin is an inhibitor of the 3c-like proteinase of severe acute respiratory syndrome coronavirus and strongly reduces virus replication in vitro.the 3c-like proteinase (3clpro) of severe acute respiratory syndrome-associated coronavirus (sars-cov) is one of the most promising targets for anti-sars-cov drugs due to its crucial role in the viral life cycle. in this study, a database containing structural information of more than 8,000 existing drugs was virtually screened by a docking approach to identify potential binding molecules of sars-cov 3clpro. as a target for screening, both a homology model and the crystallographic structure of t ...200515890949
development of a transgenic mouse model susceptible to human coronavirus 229e.human coronavirus (hcov) 229e is a group 1 coronavirus and is specific to humans. so far, no animal model is available to study the pathogenesis of infection by hcov-229e. we show here that the expression of aminopeptidase n (apn, also termed cd13), the receptor for hcov-229e, is required but not sufficient to confer susceptibility in vivo. hcov-229e infection was facilitated by crossing apn transgenic mice into signal transducers and activators of transcription (stat) 1 null mice and by adaptat ...200515919828
characterization of hcov-229e fusion core: implications for structure basis of coronavirus membrane fusion.human coronavirus 229e (hcov-229e), a member of group i coronaviruses, has been identified as one of the major viral agents causing respiratory tract diseases in humans for nearly 40 years. however, the detailed molecular mechanism of the membrane fusion mediated by the spike (s) protein of hcov-229e remains elusive. here, we report, for the first time, a rationally designed fusion core of hcov-229e (hr1-sggrgg-hr2), which was in vitro produced in gst prokaryotic expression system. multiple line ...200616714001
characterization and epitope mapping of monoclonal antibodies to the nucleocapsid protein of severe acute respiratory syndrome coronavirus.the sudden emergence of severe acute respiratory syndrome (sars) at the end of 2002 resulted in 774 reported deaths from more than 8000 cases worldwide. as no effective vaccines or antiviral agents are available, the most effective measure to prevent the expansion of a sars epidemic is the rapid diagnosis and isolation of sars patients. to establish specific diagnostic methods, we generated nine clones of monoclonal antibodies to nucleocapsid protein (np) of sars-coronavirus (sars-cov). on immun ...200818380153
the viral bronchiolites diagnosis in children by pcr multiplex.the aim of the study was to determine the etiology of the viral bronchiolites in children by using direct immunofluorescence test and 3 rt-pcr multiplex (s.bellau-pujol) the study was performed on 122 nasal inspirations collected from 3 weeks-6 month old children hospitalizated in the pediatrics service of ch rouen. the results were that the majority (53%) of bronchiolites in children had like etiology rsv and a lot of these infections had double viral etiology (26% rsv+ rhinovirus; 2,7% rsv+hmp ...200618389721
[evaluation of seeplex rv detection kit for detecting rhinovirus, human metapneumovirus, and coronavirus].direct antigen test (dat) and culture are primary tests to diagnose infections by respiratory viruses, but are mainly available for the traditional viral pathogens such as respiratory syncytial virus (rsv), influenza virus, parainfluenza virus (piv), and adenovirus in clinical laboratories. the objective of this study was to evaluate a multiplex reverse transcriptase-pcr method using seeplex rv detection kit (seegene, korea) for the detection of rhinovirus, coronavirus, and human metapneumovirus ...200818458506
crystal structures of two coronavirus adp-ribose-1''-monophosphatases and their complexes with adp-ribose: a systematic structural analysis of the viral adrp domain.the coronaviruses are a large family of plus-strand rna viruses that cause a wide variety of diseases both in humans and in other organisms. the coronaviruses are composed of three main lineages and have a complex organization of nonstructural proteins (nsp's). in the coronavirus, nsp3 resides a domain with the macroh2a-like fold and adp-ribose-1"-monophosphatase (adrp) activity, which is proposed to play a regulatory role in the replication process. however, the significance of this domain for ...200918987156
the expression and antigenicity of a truncated spike-nucleocapsid fusion protein of severe acute respiratory syndrome-associated the absence of effective drugs, controlling sars relies on the rapid identification of cases and appropriate management of the close contacts, or effective vaccines for sars. therefore, developing specific and sensitive laboratory tests for sars as well as effective vaccines are necessary for national authorities.200819038059
rapid detection and identification of 12 respiratory viruses using a dual priming oligonucleotide system-based multiplex pcr assay.acute viral respiratory infections are among the most common causes of human disease. rapid and accurate diagnosis of viral respiratory infections is important for providing timely therapeutic interventions. this study evaluated a new multiplex pcr assay (seegene inc., seoul, korea) for simultaneous detection and identification of 12 respiratory viruses using two primer mixes. the viruses included parainfluenza viruses 1, 2, and 3, human metapneumovirus, human coronavirus 229e/nl63 and oc43, ade ...200919063921
titration of human coronaviruses using an immunoperoxidase assay.calculation of infectious viral titers represents a basic and essential experimental approach for virologists. classical plaque assays cannot be used for viruses that do not cause significant cytopathic effects, which is the case for strains 229e and oc43 of human coronavirus (hcov). an alternative indirect immunoperoxidase assay (ipa) is herein described for the detection and titration of these viruses. susceptible cells are inoculated with serial logarithmic dilutions of samples in a 96-well p ...200819066576
crystal structures of the x-domains of a group-1 and a group-3 coronavirus reveal that adp-ribose-binding may not be a conserved property.the polyproteins of coronaviruses are cleaved by viral proteases into at least 15 nonstructural proteins (nsps). consisting of five domains, nsp3 is the largest of these (180-210 kda). among these domains, the so-called x-domain is believed to act as adp-ribose-1''-phosphate phosphatase or to bind poly(adp-ribose). however, here we show that the x-domain of infectious bronchitis virus (strain beaudette), a group-3 coronavirus, fails to bind adp-ribose. this is explained on the basis of the cryst ...200919177346
inhibition of human coronavirus 229e infection in human epithelial lung cells (l132) by chloroquine: involvement of p38 mapk and erk.the antiviral effects of chloroquine (cq) on human coronavirus 229e (hcov-229e) infection of human fetal lung cell line, l132 are reported. cq significantly decreased the viral replication at concentrations lower than in clinical usage. we demonstrated that cq affects the activation of p38 mitogen-activated protein kinase (mapk) and extracellular signal-regulated kinase (erk). furthermore, p38 mapk inhibitor, sb203580, inhibits cpe induced by hcov-229e infection and viral replication. our findin ...200818055026
sars-cov spike proteins expressed by the vaccinia virus tiantan strain: secreted sq protein induces robust neutralization antibody in mice.the spike (s) glycoprotein of severe acute respiratory syndrome coronavirus (sars-cov) is a major target in the development of diagnostic assays and vaccines, but its antigenic and immunogenic properties remain unclear. seven sars-cov spike proteins (s, sq, s1, rbd, s2, s2q, and cx) were generated using the modified vaccinia virus (tiantan strain) as a vector, and their antigenicity and immunogenicity were evaluated. the secreted sq protein in which the transmembrane domain was deleted, as well ...200919210229
human coronavirus and acute respiratory illness in older adults with chronic obstructive pulmonary disease.the clinical features and incidence of human coronavirus (hcov) infections in chronically ill older adults need better definition.200919239338
epidemiology and clinical presentations of human coronavirus nl63 infections in hong kong children.human coronavirus nl63 (hcov-nl63) has been found in children presenting with respiratory tract infections (rtis). however, the epidemiology and clinical course of this newly identified virus have not been fully elucidated. this study investigated the epidemiology, seasonality, and clinical features of hcov-nl63 in hong kong children. this study consisted of two cohorts of children hospitalized in a university-affiliated teaching hospital. in the 12-month retrospective part of the study, reverse ...200919759228
comprehensive detection and identification of seven animal coronaviruses and human respiratory coronavirus 229e with a microarray hybridization assay.based on microarray hybridization, a diagnostic test for coronavirus infection was developed using eight coronavirus strains: canine coronavirus (ccov), feline infectious peritonitis virus (fipv), feline coronavirus (fcov), bovine coronavirus (bcov), porcine respiratory coronavirus (prcov), turkey enteritis coronavirus (tcov), transmissible gastroenteritis virus (tgev), and human respiratory coronavirus (hrcov). up to 104 cdna clones of eight viruses were obtained by reverse transcription pcr wi ...201019955814
development of a dose-response model for sars order to develop a dose-response model for sars coronavirus (sars-cov), the pooled data sets for infection of transgenic mice susceptible to sars-cov and infection of mice with murine hepatitis virus strain 1, which may be a clinically relevant model of sars, were fit to beta-poisson and exponential models with the maximum likelihood method. the exponential model (k= 4.1 x l0(2)) could describe the dose-response relationship of the pooled data sets. the beta-poisson model did not provide a st ...201020497390
differentiation between human coronaviruses nl63 and 229e using a novel double-antibody sandwich enzyme-linked immunosorbent assay based on specific monoclonal antibodies.human coronaviruses (hcovs) are responsible for respiratory tract infections ranging from common colds to severe acute respiratory syndrome. hcov-nl63 and hcov-229e are two of the four hcovs that circulate worldwide and are close phylogenetic relatives. hcov infections can lead to hospitalization of children, elderly individuals, and immunocompromised patients. globally, approximately 5% of all upper and lower respiratory tract infections in hospitalized children are caused by hcov-229e and hcov ...201021084464
glucose-6-phosphate dehydrogenase deficiency enhances human coronavirus 229e infection.the host cellular environment is a key determinant of pathogen infectivity. viral gene expression and viral particle production of glucose-6-phosphate dehydrogenase (g6pd)-deficient and g6pd-knockdown cells were much higher than their counterparts when human coronavirus (hcov) 229e was applied at 0.1 multiplicity of infection. these phenomena were correlated with increased oxidant production. accordingly, ectopic expression of g6pd in g6pd-deficient cells or addition of antioxidant (such as alph ...200818269318
outbreaks of human coronavirus in a pediatric and neonatal intensive care unit.human coronavirus 229e (hcov) has been recently recognized as a potential agent of nosocomial viral respiratory infections (nrvi) in high-risk infants. we have confirmed this as fact through the study of a 1-year period of hcov outbreaks occurring during a prospective survey of nrvi in a paediatric and neonatal intensive care unit (pnicu) using new molecular techniques for hcov detection. nasal samples obtained at admission and weekly thereafter for all hospitalised children, as well as monthly ...200818335238
human coronavirus nl63 employs the severe acute respiratory syndrome coronavirus receptor for cellular entry.coronavirus (cov) infection of humans is usually not associated with severe disease. however, discovery of the severe acute respiratory syndrome (sars) cov revealed that highly pathogenic human covs (hcovs) can evolve. the identification and characterization of new hcovs is, therefore, an important task. recently, a hcov termed nl63 was discovered in patients with respiratory tract illness. here, cell tropism and receptor usage of hcov-nl63 were analyzed. the nl63 spike (s) protein mediated infe ...200515897467
antigenic cross-reactivity between severe acute respiratory syndrome-associated coronavirus and human coronaviruses 229e and oc43.cross-reactivity between antibodies to different human coronaviruses (hcovs) has not been systematically studied. by use of western blot analysis, indirect immunofluorescence assay (ifa), and enzyme-linked immunosorbent assay (elisa), antigenic cross-reactivity between severe acute respiratory syndrome (sars)-associated coronavirus (sars-cov) and 2 hcovs (229e and oc43) was demonstrated in immunized animals and human serum. in 5 of 11 and 10 of 11 patients with sars, paired serum samples showed ...200515897988
human coronavirus 229e remains infectious on common touch surface materials.the evolution of new and reemerging historic virulent strains of respiratory viruses from animal reservoirs is a significant threat to human health. inefficient human-to-human transmission of zoonotic strains may initially limit the spread of transmission, but an infection may be contracted by touching contaminated surfaces. enveloped viruses are often susceptible to environmental stresses, but the human coronaviruses responsible for severe acute respiratory syndrome (sars) and middle east respi ...201526556276
evidence for an ancestral association of human coronavirus 229e with bats.we previously showed that close relatives of human coronavirus 229e (hcov-229e) exist in african bats. the small sample and limited genomic characterizations have prevented further analyses so far. here, we tested 2,087 fecal specimens from 11 bat species sampled in ghana for hcov-229e-related viruses by reverse transcription-pcr (rt-pcr). only hipposiderid bats tested positive. to compare the genetic diversity of bat viruses and hcov-229e, we tested historical isolates and diagnostic specimens ...201526378164
comparison of the luminex xtag respiratory viral panel with xtag respiratory viral panel fast for diagnosis of respiratory virus infections.nucleic acid tests are sensitive and specific and provide a rapid diagnosis, making them invaluable for patient and outbreak management. multiplex pcr assays have additional advantages in providing an economical and comprehensive panel for many common respiratory viruses. previous reports have shown the utility of the xtag respiratory viral panel (rvp) assay manufactured by luminex molecular diagnostics for this purpose. a newer generation of this kit, released in canada in early 2010, is design ...201121411570
blocking eif4e:eif4g interaction as a strategy to impair coronavirus replication.coronaviruses are a family of enveloped single-stranded positive-sense rna viruses causing respiratory, enteric and neurologic diseases in mammals and fowl. human coronaviruses are recognized to cause up to a third of common colds and are suspected to be involved in enteric and neurologic diseases. coronavirus replication involves generation of nested subgenomic (sg) mrnas with a common capped 5' leader sequence. translation of most of the sgmrnas is thought to be cap-dependent and displays a re ...201121507972
identification and characterization of a novel alpaca respiratory coronavirus most closely related to the human coronavirus 2007, a novel coronavirus associated with an acute respiratory disease in alpacas (alpaca coronavirus, acov) was isolated. full-length genomic sequencing of the acov demonstrated the genome to be consistent with other alphacoronaviruses. a putative additional open-reading frame was identified between the nucleocapsid gene and 3'utr. the acov was genetically most similar to the common human coronavirus (hcov) 229e with 92.2% nucleotide identity over the entire genome. a comparison of spike gen ...201223235471
bat origin of human coronaviruses.bats have been recognized as the natural reservoirs of a large variety of viruses. special attention has been paid to bat coronaviruses as the two emerging coronaviruses which have caused unexpected human disease outbreaks in the 21st century, severe acute respiratory syndrome coronavirus (sars-cov) and middle east respiratory syndrome coronavirus (mers-cov), are suggested to be originated from bats. various species of horseshoe bats in china have been found to harbor genetically diverse sars-li ...201526689940
neurological manifestations of covid-19 and other coronaviruses: a systematic describe the main neurological manifestations related to coronavirus infection in humans.202032834527
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