Publications

severe acute respiratory syndrome coronavirus infection in children.		2005	16118891
[a preliminary study of the structure prediction and expression of sars-cov spike protein].	in this study, the encoding sequences of sars-cov spike protein were analyzed by bioinformatics methods, the structural characteristics and functions were forecasted based on available data. it suggests that the fragment of spike glycoprotein (s401-659) may be crucial for viral attachment and may be a major immunodominant epitope. then the fragment was amplified and subcloned into expression vector pet28a(+) and ppic9k. these two plasmids pet28a(+)-s and ppic9k-s were transformed to e.coli strai ...	2005	16120590
a serological survey on neutralizing antibody titer of sars convalescent sera.	a seroepidemiologic study was conducted in north china in 2003 to determine the neutralizing antibody titer of severe acute respiratory syndrome (sars) convalescent sera. a total of 99 sars convalescent serum samples were collected from patients from the inner mongolia autonomous region, hebei province, and beijing 35-180 days after the onset of symptoms. the anti-sars antibodies were detected by enzyme-linked immunosorbent assay (elisa), neutralization assay, and western blot. eighty-seven seru ...	2005	16121363
rapid and sensitive detection of multiple genes from the sars-coronavirus using quantitative rt-pcr with dual systems.	the outbreak of severe acute respiratory syndrome (sars) was caused by a newly identified coronavirus (sars-cov) in 2003. to detect early sars-cov infection, a one-step, real-time quantitative reverse transcription-polymerase chain reaction (rt-pcr) assay was developed that could simultaneously detect nucleocapsid (n), membrane (m), and spike (s) genes of sars-cov with the same pcr condition using either applied biosystems (abi) prism 7700 sequence detection system or roche lightcycler. the sens ...	2005	16121372
respiratory disease caused by a species b2 adenovirus in a military camp in turkey.	in april 2004, two patients were admitted to hospital in berlin, germany, with clinical signs of acute respiratory infection after returning from a military exercise in their home country of turkey. they were admitted to a high security infectious disease unit as epidemiological data pointed to an outbreak of unknown etiology. samples taken at the time of admission proved to be strongly positive for adenovirus by pcr, but negative for influenza a/h1n1 virus, influenza a/h3n2 virus, influenza b v ...	2005	16121380
the sars coronavirus s glycoprotein receptor binding domain: fine mapping and functional characterization.	the entry of the sars coronavirus (scv) into cells is initiated by binding of its spike envelope glycoprotein (s) to a receptor, ace2. we and others identified the receptor-binding domain (rbd) by using s fragments of various lengths but all including the amino acid residue 318 and two other potential glycosylation sites. to further characterize the role of glycosylation and identify residues important for its function as an interacting partner of ace2, we have cloned, expressed and characterize ...	2005	16122388
recent highlights in the development of new antiviral drugs.	twenty antiviral drugs, that is about half of those that are currently approved, are formally licensed for clinical use in the treatment of human immunodeficiency virus infections (acquired immune deficiency syndrome). the others are used in the treatment of herpesvirus (e.g. herpes simplex virus, varicella zoster virus and cytomegalo virus), hepatitis b virus, hepatitis c virus or influenza virus infections. recent endeavours have focussed on the development of improved antiviral therapies for ...	2005	16125443
design of wide-spectrum inhibitors targeting coronavirus main proteases.	the genus coronavirus contains about 25 species of coronaviruses (covs), which are important pathogens causing highly prevalent diseases and often severe or fatal in humans and animals. no licensed specific drugs are available to prevent their infection. different host receptors for cellular entry, poorly conserved structural proteins (antigens), and the high mutation and recombination rates of covs pose a significant problem in the development of wide-spectrum anti-cov drugs and vaccines. cov m ...	2005	16128623
rational monoclonal antibody development to emerging pathogens, biothreat agents and agents of foreign animal disease: the antigen scale.	many factors influence the choice of methods used to develop antibody to infectious agents. in this paper, we review the current status of the main technologies used to produce monoclonal antibodies (mabs) from the b cells of antigen-sensitized animals. while companies are adopting advanced high-throughput methods, the major technologies used by veterinary and medical research laboratories are classical hybridoma fusion and recombinant library selection techniques. these methods have inherent ad ...	2005	16129340
efficient and quick inactivation of sars coronavirus and other microbes exposed to the surfaces of some metal catalysts.	to study the two metal catalysts ag/al2o3 and cu/al2o3 that interdict the transmission pathway for sars and other respiratory infectious diseases.	2005	16131020
selective immobilization of fusion proteins on poly(hydroxyalkanoate) microbeads.	a novel fusion protein system employing the substrate-binding domain (sbd) of poly(hydroxyalkanoate) (pha) depolymerase was developed for the specific immobilization of proteins on pha microbeads, and was consequently used for immunoassays. the enhanced green fluorescent protein, red fluorescent protein, and severe acute respiratory syndrome coronavirus envelope protein were used as model proteins, and were selectively and functionally immobilized to the pha microbeads by fusing them to the sbd. ...	2005	16131092
characterization of neutralizing monoclonal antibodies recognizing a 15-residues epitope on the spike protein hr2 region of severe acute respiratory syndrome coronavirus (sars-cov).	the spike (s) glycoprotein is thought to play a complex and central role in the biology and pathogenesis of sars coronavirus infection. in this study, a recombinant protein (rs268, corresponding to residues 268-1255 of sars-cov s protein) was expressed in escherichia coli and was purified to near homogeneity. after immunization with rs268, s protein-specific balb/c antisera and mabs were induced and confirmed using elisa, western blot and ifa. several balb/c mabs were found to be effectively to ...	2005	16132115
double-antigen sandwich elisa for detection of antibodies to sars-associated coronavirus in human serum.	the study presented here was conducted to evaluate the performance of a double-antigen sandwich elisa to detect antibodies in human serum against the coronavirus associated with severe acute respiratory syndrome (sars). a recombinant partial nucleocapsid protein of sars-associated coronavirus was used as a serodiagnostic antigen in the elisa. a total of 2892 clinical serum samples were tested with the elisa kit, which positively identified 25 of 35 (71.4%) samples of patients with confirmed sars ...	2005	16133409
selection of and recombination between minor variants lead to the adaptation of an avian coronavirus to primate cells.	an interesting question posed by the current evidence that severe acute respiratory syndrome coronavirus may be originated from an animal coronavirus is how such an animal coronavirus breaks the host species barrier and becomes zoonotic. in this report, we study the chronological order of genotypic changes in the spike protein of avian coronavirus infectious bronchitis virus (ibv) during its adaptation to a primate cell line. adaptation of the beaudette strain of ibv from chicken embryo to vero ...	2005	16137658
selection of and recombination between minor variants lead to the adaptation of an avian coronavirus to primate cells.	an interesting question posed by the current evidence that severe acute respiratory syndrome coronavirus may be originated from an animal coronavirus is how such an animal coronavirus breaks the host species barrier and becomes zoonotic. in this report, we study the chronological order of genotypic changes in the spike protein of avian coronavirus infectious bronchitis virus (ibv) during its adaptation to a primate cell line. adaptation of the beaudette strain of ibv from chicken embryo to vero ...	2005	16137658
single amino acid substitutions in the severe acute respiratory syndrome coronavirus spike glycoprotein determine viral entry and immunogenicity of a major neutralizing domain.	neutralizing antibodies (nabs) against severe acute respiratory syndrome (sars) coronavirus (sars-cov) spike (s) glycoprotein confer protection to animals experimentally infected with the pathogenic virus. we and others previously demonstrated that a major mechanism for neutralizing sars-cov was through blocking the interaction between the s glycoprotein and the cellular receptor angiotensin-converting enzyme 2 (ace2). in this study, we used in vivo electroporation dna immunization and a pseudov ...	2005	16140741
molecular evolution analysis and geographic investigation of severe acute respiratory syndrome coronavirus-like virus in palm civets at an animal market and on farms.	massive numbers of palm civets were culled to remove sources for the reemergence of severe acute respiratory syndrome (sars) in guangdong province, china, in january 2004, following sars coronavirus detection in market animals. the virus was identified in all 91 palm civets and 15 raccoon dogs of animal market origin sampled prior to culling, but not in 1,107 palm civets later sampled at 25 farms, spread over 12 provinces, which were claimed to be the source of traded animals. twenty-seven novel ...	2005	16140765
molecular evolution analysis and geographic investigation of severe acute respiratory syndrome coronavirus-like virus in palm civets at an animal market and on farms.	massive numbers of palm civets were culled to remove sources for the reemergence of severe acute respiratory syndrome (sars) in guangdong province, china, in january 2004, following sars coronavirus detection in market animals. the virus was identified in all 91 palm civets and 15 raccoon dogs of animal market origin sampled prior to culling, but not in 1,107 palm civets later sampled at 25 farms, spread over 12 provinces, which were claimed to be the source of traded animals. twenty-seven novel ...	2005	16140765
survival of severe acute respiratory syndrome coronavirus.	the primary modes of transmission of severe acute respiratory syndrome (sars) coronavirus (sars-cov) appear to be direct mucus membrane contact with infectious droplets and through exposure to formites. knowledge of the survival characteristics of the virus is essential for formulating appropriate infection-control measures.	2005	16142653
activation of nf-kappab by the full-length nucleocapsid protein of the sars coronavirus.	the severe acute respiratory syndrome coronavirus (sars-cov) is the major causative agent for the worldwide outbreak of sars in 2003. the mechanism by which sars-cov causes atypical pneumonia remains unclear. the nuclear factor kappa b (nf-kappab) is a key transcription factor that activates numerous genes involved in cellular immune response and inflammation. many studies have shown that nf-kappab plays an important role in the pathogenesis of lung diseases. in this study, we investigated the p ...	2005	16143815
sars-cov genome polymorphism: a bioinformatics study.	a dataset of 103 sars-cov isolates (101 human patients and 2 palm civets) was investigated on different aspects of genome polymorphism and isolate classification. the number and the distribution of single nucleotide variations (snvs) and insertions and deletions, with respect to a "profile", were determined and discussed ("profile" being a sequence containing the most represented letter per position). distribution of substitution categories per codon positions, as well as synonymous and non-syno ...	2005	16144519
optimal word sizes for dissimilarity measures and estimation of the degree of dissimilarity between dna sequences.	several measures of dna sequence dissimilarity have been developed. the purpose of this paper is 3-fold. firstly, we compare the performance of several word-based or alignment-based methods. secondly, we give a general guideline for choosing the window size and determining the optimal word sizes for several word-based measures at different window sizes. thirdly, we use a large-scale simulation method to simulate data from the distribution of sk-ld (symmetric kullback-leibler discrepancy). these ...	2005	16144805
protection of mammalian cells from severe acute respiratory syndrome coronavirus infection by equine neutralizing antibody.	the aetiological agent for severe acute respiratory syndrome (sars) has been determined to be a new type of coronavirus (sars-cov) that infects a wide range of mammalian hosts. up to now, there have been no specific drugs to protect against sars-cov infection, thus developing effective strategies against this newly emerged viral infection warrants urgent efforts. adoptive immune therapy with pathogen-specific heterologous immunoglobulin has been successfully used to control the dissemination of ...	2005	16152762
screening and identification of linear b-cell epitopes and entry-blocking peptide of severe acute respiratory syndrome (sars)-associated coronavirus using synthetic overlapping peptide library.	a 10-mer overlapping peptide library has been synthesized for screening and identification of linear b-cell epitopes of severe acute respiratory syndrome associated coronavirus (sars-cov), which spanned the major structural proteins of sars-cov. one hundred and eleven candidate peptides were positive according to the result of pepscan, which were assembled into 22 longer peptides. five of these peptides showed high cross-immunoreactivities (approximately 66.7 to 90.5%) to sars convalescent patie ...	2005	16153058
detection of the nucleocapsid protein of severe acute respiratory syndrome coronavirus in serum: comparison with results of other viral markers.	a capture enzyme-enhanced chemiluminescence immunoassay (eclia) based on three specific monoclonal antibodies to detect the nucleocapsid (n) protein of severe acute respiratory syndrome (sars) associated coronavirus (sars-cov) in the serial serum samples from sars patients was developed. the anti-sars-cov igg and the viral rna were also detected in the sera by elisa and rt-pcr, respectively. during the first 10 days after onset, anti-sars-cov igg, sars-cov rna and the n protein were detected in ...	2005	16024098
[detection of autoimmune parameter of sars patients].	to explore whether autoimmune phenomena exist in sars patients, and to seek for unusual autoimmune antibodies in sars patients.	2005	16027775
[conditions for sars-cov cultivation and inactivation].	to study the method for cultivation and inactivation of sars-cov.	2005	16027779
[determination of sars-cov by simplified nested fluorescent rt-pcr].	to establish a simple rapid and sensitive nested rt-pcr method for detection of sars coronavirus rna by designing the specific primers for sars and optimizing the parameters for pcr.	2005	16027791
autoantibodies against human epithelial cells and endothelial cells after severe acute respiratory syndrome (sars)-associated coronavirus infection.	the severe acute respiratory syndrome (sars) is caused by infection with the sars-associated coronavirus (sars-cov) and characterized by severe pulmonary inflammation and fibrosis. in this study, the development of autoantibodies against human epithelial cells and endothelial cells in patients with sars at different time periods (the first week: phase i, 1 month after the disease onset: phase ii/phase iii) were investigated. antibodies in sera of patients and healthy controls against: (1) a549 h ...	2005	16032747
vesicular stomatitis virus pseudotyped with severe acute respiratory syndrome coronavirus spike protein.	severe acute respiratory syndrome coronavirus (sars-cov) contains a single spike (s) protein, which binds to its receptor, angiotensin-converting enzyme 2 (ace2), induces membrane fusion and serves as a neutralizing antigen. a sars-cov-s protein-bearing vesicular stomatitis virus (vsv) pseudotype using the vsvdeltag* system was generated. partial deletion of the sars-cov-s protein cytoplasmic domain allowed efficient incorporation into vsv particles and led to the generation of a pseudotype (vsv ...	2005	16033974
vaccine design for severe acute respiratory syndrome coronavirus.	severe acute respiratory syndrome (sars) is an emerging infectious disease caused by a new coronavirus (sars-cov). recent studies suggest that sars-cov is zoonotic and may have a broad host range besides humans. although the global outbreak of sars has been contained, there are serious concerns over its re-emergence and bioterrorism potential. as a part of preparedness, development of a safe and effective vaccine is one of the highest priorities in fighting sars. a number of candidate vaccines, ...	2005	16035944
excretion and detection of sars coronavirus and its nucleic acid from digestive system.	to study whether severe acute respiratory syndrome coronavirus (sars-cov) could be excreted from digestive system.	2005	16038039
inhibition of replication and infection of severe acute respiratory syndrome-associated coronavirus with plasmid-mediated interference rna.	severe acute respiratory syndrome (sars) is a newly emerged infectious disease caused by a novel coronavirus (sars-cov), which spread to over 30 countries in early 2003. until recently, no specific vaccines and effective drugs have been available to protect patients from infection by this virus. to exploit a new strategy to fight this disease, we investigated the effect of interference rna (rnai) on the virus infection and replication with 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bro ...	2005	16038478
high-resolution structure of hla-a*1101 in complex with sars nucleocapsid peptide.	the structure of the human mhc-i molecule hla-a1101 in complex with a nonameric peptide (ktfpptepk) has been determined by x-ray crystallography to 1.45 a resolution. the peptide is derived from the sars-cov nucleocapsid protein positions 362-370 (snp362-370). it is conserved in all known isolates of sars-cov and has been verified by in vitro peptide-binding studies to be a good to intermediate binder to hla-a0301 and hla-a*1101, with ic50 values of 70 and 186 nm, respectively [sylvester-hvid ...	2005	16041067
papain-like protease 2 (plp2) from severe acute respiratory syndrome coronavirus (sars-cov): expression, purification, characterization, and inhibition.	viral proteases are essential for pathogenesis and virulence of severe acute respiratory syndrome coronavirus (sars-cov). little information is available on sars-cov papain-like protease 2 (plp2), and development of inhibitors against plp2 is attractive for antiviral therapy. here, we report the characterization of sars-cov plp2 (from residues 1414 to 1858) purified from baculovirus-infected insect cells. we demonstrate that sars-cov plp2 by itself differentially cleaves between the amino acids ...	2005	16042412
long-term protection from sars coronavirus infection conferred by a single immunization with an attenuated vsv-based vaccine.	although the recent sars coronavirus (sars-cov) that appeared in 2002 has now been contained, the possibility of re-emergence of sars-cov remains. due to the threat of re-emergence, the overall fatality rate of approximately 10%, and the rapid dispersion of the virus via international travel, viable vaccine candidates providing protection from sars are clearly needed. we developed an attenuated vsv recombinant (vsv-s) expressing the sars coronavirus (sars-cov) spike (s) protein. in cells infecte ...	2005	16043204
multiple organ infection and the pathogenesis of sars.	after >8,000 infections and >700 deaths worldwide, the pathogenesis of the new infectious disease, severe acute respiratory syndrome (sars), remains poorly understood. we investigated 18 autopsies of patients who had suspected sars; 8 cases were confirmed as sars. we evaluated white blood cells from 22 confirmed sars patients at various stages of the disease. t lymphocyte counts in 65 confirmed and 35 misdiagnosed sars cases also were analyzed retrospectively. sars viral particles and genomic se ...	2005	16043521
antibody to severe acute respiratory syndrome (sars)-associated coronavirus spike protein domain 2 cross-reacts with lung epithelial cells and causes cytotoxicity.	both viral effect and immune-mediated mechanism are involved in the pathogenesis of severe acute respiratory syndrome-associated coronavirus (sars-cov) infection. in this study, we showed that in sars patient sera there were autoantibodies (autoabs) that reacted with a549 cells, the type-2 pneumocytes, and that these autoabs were mainly igg. the autoabs were detectable 20 days after fever onset. tests of non-sars-pneumonia patients did not show the same autoab production as in sars patients. aft ...	2005	16045740
proposal for vaccination against sars coronavirus using avian infectious bronchitis virus strain h from the netherlands.		2005	16045996
nucleolar localization of non-structural protein 3b, a protein specifically encoded by the severe acute respiratory syndrome coronavirus.	the open reading frame 3 (orf3) of the severe acute respiratory syndrome coronavirus (sars-cov) genome encodes a predicted 154-amino acid protein, which lacks similarities to any known protein, and is named 3b. in this study, it was shown that 3b protein was predominately localized to nucleus with egfp tag at its n- or c-terminus. the localization patterns were similar in different transfected cells. immuno-fluorescence assay revealed that 3b protein was co-localized well with c23 in nucleolus. ...	2005	16046244
bcl-xl inhibits t-cell apoptosis induced by expression of sars coronavirus e protein in the absence of growth factors.	one of the hallmark findings in patients suffering from sars (severe acute respiratory syndrome) is lymphopenia, which is the result of massive lymphocyte death. sars-cov (sars coronavirus), a novel coronavirus that has been etiologically associated with sars cases, is homologous with mhv (murine hepatitis coronavirus), and mhv small envelope e protein is capable of inducing apoptosis. we hypothesized that sars-cov encodes a small envelope e protein that is homologous with mhv e protein, thus in ...	2005	16048439
pneumonitis and multi-organ system disease in common marmosets (callithrix jacchus) infected with the severe acute respiratory syndrome-associated coronavirus.	severe acute respiratory syndrome (sars) is a significant emerging infectious disease. humans infected with the etiological agent, sars-associated coronavirus (sars-cov), primarily present with pneumonitis but may also develop hepatic, gastrointestinal, and renal pathology. we inoculated common marmosets (callithrix jacchus) with the objective of developing a small nonhuman primate model of sars. two groups of c. jacchus were inoculated intratracheally with cell culture supernatant containing sa ...	2005	16049331
emerging infectious agents.	as new agents of infectious disease continue to emerge and old antagonists reemerge, it is clear that the war on infectious disease is far from over. indeed, the appearance of sars during the past year is the latest example of the continuing challenges posed by agents of infectious disease. emerging agents are threats not only to the general population, but also to recipients of blood transfusions. today a variety of emerging agents are of concern to transfusion safety including trypanosoma cruz ...	2005	16050150
lsectin interacts with filovirus glycoproteins and the spike protein of sars coronavirus.	cellular attachment factors like the c-type lectins dc-sign and dc-signr (collectively referred to as dc-sign/r) can augment viral infection and might promote viral dissemination in and between hosts. the lectin lsectin is encoded in the same chromosomal locus as dc-sign/r and is coexpressed with dc-signr on sinusoidal endothelial cells in liver and lymphnodes. here, we show that lsectin enhances infection driven by filovirus glycoproteins (gp) and the s protein of sars coronavirus, but does not ...	2005	16051304
peptide domain involved in the interaction between membrane protein and nucleocapsid protein of sars-associated coronavirus.	severe acute respiratory syndrome (sars) is an emerging infectious disease associated with a novel coronavirus (cov) that was identified and molecularly characterized in 2003. previous studies on various coronaviruses indicate that protein-protein interactions amongst various coronavirus proteins are critical for viral assembly and morphogenesis. it is necessary to elucidate the molecular mechanism of sars-cov replication and rationalize the anti-sars therapeutic intervention. in this study, we ...	2005	16053703
the severe acute respiratory syndrome (sars)-coronavirus 3a protein may function as a modulator of the trafficking properties of the spike protein.	background: a recent publication reported that a tyrosine-dependent sorting signal, present in cytoplasmic tail of the spike protein of most coronaviruses, mediates the intracellular retention of the spike protein. this motif is missing from the spike protein of the severe acute respiratory syndrome-coronavirus (sars-cov), resulting in high level of surface expression of the spike protein when it is expressed on its own in vitro. presentation of the hypothesis: it has been shown that the severe ...	2005	15703085
reference gene selection for quantitative real-time pcr analysis in virus infected cells: sars corona virus, yellow fever virus, human herpesvirus-6, camelpox virus and cytomegalovirus infections.	ten potential reference genes were compared for their use in experiments investigating cellular mrna expression of virus infected cells. human cell lines were infected with cytomegalovirus, human herpesvirus-6, camelpox virus, sars coronavirus or yellow fever virus. the expression levels of these genes and the viral replication were determined by real-time pcr. genes were ranked by the bestkeeper tool, the genorm tool and by criteria we reported previously. ranking lists of the genes tested were ...	2005	15705200
border screening for sars.	with the rapid international spread of severe acute respiratory syndrome (sars) from march through may 2003, canada introduced various measures to screen airplane passengers at selected airports for symptoms and signs of sars. the world health organization requested that all affected areas screen departing passengers for sars symptoms. in spite of intensive screening, no sars cases were detected. sars has an extremely low prevalence, and the positive predictive value of screening is essentially ...	2005	15705315
anti-sars-cov immunoglobulin g in healthcare workers, guangzhou, china.	to determine the prevalence of inapparent infection with severe acute respiratory syndrome (sars) among healthcare workers, we performed a serosurvey to test for immunoglobulin (ig) g antibodies to the sars coronavirus (sars-cov) among 1,147 healthcare workers in 3 hospitals that admitted sars patients in mid-may 2003. among them were 90 healthcare workers with sars. as a reference group, 709 healthcare workers who worked in 2 hospitals that never admitted any sars patients were similarly tested ...	2005	15705328
sars clinical features, united states, 2003.	we compared the clinical features of 8 u.s. case-patients with laboratory-confirmed severe acute respiratory syndrome (sars) to 65 controls who tested negative for sars coronavirus (sars-cov) infection. shortness of breath, vomiting, diarrhea, progressive bilateral infiltrates on chest radiograph, and need for supplemental oxygen were significantly associated with confirmed sars-cov infection.	2005	15705339
new antiviral drugs, vaccines and classic public health interventions against sars coronavirus.	severe acute respiratory syndrome (sars) is caused by one of two recently discovered coronaviruses. the virus is emergent from south east (se) asian mammals: either the civet cat, a related species or a rat species. the virus has a long incubation period and low reproduction number (r0 value) and hence the first outbreak in 2004 was controlled by hygiene and quarantine. however, the healthcare system was compromised and the economic cost was extremely high. fortunately, the virus is easily culti ...	2005	15739618
current status of anti-sars agents.	severe acute respiratory syndrome (sars) is a disease that has newly emerged in the 21st century, and is both severe and highly contagious. sars first surfaced in late 2002 and spread within a few months from its origin in guandong province, china, to more than 30 countries (world health organization, 2003). in this review, several antiviral substances shown to be active in vitro will be introduced and summarized in the order of the virus' replication steps; that is, binding to cellular receptor ...	2005	15739619
the protein x4 of severe acute respiratory syndrome-associated coronavirus is expressed on both virus-infected cells and lung tissue of severe acute respiratory syndrome patients and inhibits growth of balb/c 3t3 cell line.	the genome of the severe acute respiratory syndrome-associated coronavirus (sars-cov) includes sequences encoding the putative protein x4 (orf8, orf7a), consisting of 122 amino acids. the deduced sequence contains a probable cleaved signal peptide sequence and a c-terminal transmembrane helix, indicating that protein x4 is likely to be a type i membrane protein. this study was conducted to demonstrate whether the protein x4 was expressed and its essential function in the process of sars-cov infe ...	2005	15740663
[specific antibodies against the nucleocapsid protein and s1 domain of spike glycoprotein in patients with severe acute respiratory syndrome].	to explore the laws of the appearance of the specific serum antibodies against the nucleocapsid (n) protein and the s1 domain of spike (s) glycoprotein in patients with severe acute respiratory syndrome (sars) and to evaluate the value of these two proteins to be used as diagnostic makers for sars.	2005	15949331
[an immunofluorescence assay for the detection of sars associated coronavirus antibody based on recombinant nucleocapsid antigen and its application].	to establish a new technique for sars-cov antibody test to detect infection of severer acute respiratory syndrome (sars).	2005	15949361
screening of drugs by fret analysis identifies inhibitors of sars-cov 3cl protease.	sars-cov 3cl protease is essential for viral protein processing and is regarded as a good drug target to prevent sars-cov replication. in the present study, we established a high-throughput fret technique for screening for anti-sars-cov 3cl protease drugs. of a thousand existing drugs examined, hexachlorophene was identified as the most potent in inhibiting sars-cov 3cl protease. further characterization showed that it was effective at micromolar concentrations (k(i) = 4 microm). the binding mod ...	2005	15950190
pathogenesis of severe acute respiratory syndrome.	severe acute respiratory syndrome (sars) is a zoonotic infectious disease caused by a novel coronavirus (cov). the tissue tropism of sars-cov includes not only the lung, but also the gastrointestinal tract, kidney and liver. angiotensin-converting enzyme 2 (ace2), the c-type lectin cd209l (also known l-sign), and dc-sign bind sars-cov, but ace2 appears to be the key functional receptor for the virus. there is a prominent innate immune response to sars-cov infection, including acute-phase protein ...	2005	15950449
multiplexed detection of pathogen dna with dna-based fluorescence nanobarcodes.	rapid, multiplexed, sensitive and specific molecular detection is of great demand in gene profiling, drug screening, clinical diagnostics and environmental analysis. one of the major challenges in multiplexed analysis is to identify each specific reaction with a distinct label or 'code'. two encoding strategies are currently used: positional encoding, in which every potential reaction is preassigned a particular position on a solid-phase support such as a dna microarray, and reaction encoding, w ...	2005	15951805
severe acute respiratory syndrome (sars) s protein production in plants: development of recombinant vaccine.	in view of a recent spread of severe acute respiratory syndrome (sars), there is a high demand for production of a vaccine to prevent this disease. recent studies indicate that sars-coronavirus (cov) spike protein (s protein) and its truncated fragments are considered the best candidates for generation of the recombinant vaccine. toward the development of a safe, effective, and inexpensive vaccine candidate, we have expressed the n-terminal fragment of sars-cov s protein (s1) in tomato and low-n ...	2005	15956182
the 3a protein of severe acute respiratory syndrome-associated coronavirus induces apoptosis in vero e6 cells.	an outbreak of severe acute respiratory syndrome (sars) occurred in china and the first case emerged in mid-november 2002. the aetiological agent of this disease was found to be a previously unknown coronavirus, sars-associated coronavirus (sars-cov). the detailed pathology of sars-cov infection and the host response to the viral infection are still not known. the 3a gene encodes a non-structural viral protein, which is predicted to be a transmembrane protein. in this study, it was shown that th ...	2005	15958670
development of a homogeneous screening assay for automated detection of antiviral agents active against severe acute respiratory syndrome-associated coronavirus.	the severity and global spread of the 2003 outbreak of the severe acute respiratory syndrome-associated coronavirus (sars-cov) highlighted the risks to human health posed by emerging viral diseases and emphasized the need for specific therapeutic agents instead of relying on existing broadly active antiviral compounds. the development of rapid screening assays is essential for antiviral drug discovery. thus, a screening system for anti-sars-cov agents was developed, which evaluated compound pote ...	2005	15961169
appropriate use of personal protective equipment among healthcare workers in public sector hospitals and primary healthcare polyclinics during the sars outbreak in singapore.	singapore was affected by an outbreak of severe acute respiratory syndrome (sars) from 25 february to 31 may 2003, with 238 probable cases and 33 deaths.	2005	15961624
genetic lesions within the 3a gene of sars-cov.	a series of frameshift mutations within the 3a gene has been observed in culture-derived severe acute respiratory syndrome coronavirus (sars-cov). we report here that viral rna from clinical samples obtained from sars-cov infected patients also contains a heterogeneous population of wild-type and mutant 3a transcripts.	2005	15963240
concentration and detection of sars coronavirus in sewage from xiao tang shan hospital and the 309th hospital.	the transmission of severe acute respiratory syndrome-associated coronavirus (sars-cov) is associated with close contact to sars patients and droplet secretions of those patients. the finding of positive rt-pcr results from stools of sars patients suggests that stools of sars patients or sewage containing stools of patients could transmit sars-cov. we used a novel style of electropositive filter media particle to concentrate the sars-cov from the sewage of two hospitals receiving sars patients i ...	2005	15964082
longitudinal alteration of circulating dendritic cell subsets and its correlation with steroid treatment in patients with severe acute respiratory syndrome.	in this study, we found that 74 patients with severe acute respiratory syndrome (sars) exhibited a rapid, dramatic decrease in numbers of circulating myeloid and plasmacytoid dendritic cells (mdcs and pdcs) during the first 2 weeks of illness (5.3- and 28.4-fold reductions for mdcs and pdcs compared with 25 healthy individuals, respectively), with slow return to normal cell numbers during convalescence (weeks 5-7 of illness on average). in addition, numbers of circulating cd4 and cd8 t cells exh ...	2005	15964242
purification of severe acute respiratory syndrome hyperimmune globulins for intravenous injection from convalescent plasma.	severe acute respiratory syndrome (sars) is a new infectious disease caused by the sars virus. current first-line treatments are experimental, and their effectiveness remains open to question. for more effective treatment and prevention of sars, human sars hyperimmune globulins for intravenous (iv) injection were purified in this study.	2005	15987362
immune response in balb/c mice induced by recombinant spike protein of severe acute respiratory syndrome coronavirus.		2005	15989769
[construction of a large human scfv library against sars virus].	to construct a large human scfv library against sars virus by using in vivo recombination.	2005	15989788
susceptibility of human and rat neural cell lines to infection by sars-coronavirus.	pathological characterization of autopsied tissues from patients with sars revealed severe damage in restricted tissues, such as lung, with no apparent cell damage in other tissues, such as intestine and brain. here, we examined the susceptibility of neural cell lines of human (ol) and rat (c6) origins to sars-associated coronavirus. both of the neural cell lines showed no apparent cytopathic effects (cpe) by infection but produced virus with infectivity of 10(2-5) per ml, in sharp contrast to t ...	2005	15992768
nuclear/nucleolar localization properties of c-terminal nucleocapsid protein of sars coronavirus.	a novel coronavirus (cov) has recently been identified as the aetiological agent of severe acute respiratory syndrome (sars). nucleocapsid (n) proteins of the coronaviridae family have no discernable homology, but they share a common nucleolar-cytoplasmic distribution pattern. there are three putative nuclear localization signal (nls) motifs present in the n. to determine the role of these putative nlss in the intracellular localization of the sars-cov n, we performed a confocal microscopy analy ...	2005	15992957
reduced bone mineral density in male severe acute respiratory syndrome (sars) patients in hong kong.	during the severe acute respiratory syndrome (sars) outbreak in hong kong in 2003, patients were treated with very high doses of corticosteroid and ribavirin. the detrimental effects of such treatment on the bone mineral density (bmd) of sars patients are unknown. to compare the bmd of sars patients with normal range data, a cross-sectional survey was conducted. the bone mineral density of 224 patients with sars, who were treated with an average of 2753 mg (sd = 2152 mg) prednisolone and 29,344 ...	2005	15993669
sars coronavirus spike polypeptide dna vaccine priming with recombinant spike polypeptide from escherichia coli as booster induces high titer of neutralizing antibody against sars coronavirus.	different forms of sars coronavirus (sars-cov) spike protein-based vaccines for generation of neutralizing antibody response against sars-cov were compared using a mouse model. high igg levels were detected in mice immunized with intraperitoneal (i.p.) recombinant spike polypeptide generated by escherichia coli (s-peptide), mice primed with intramuscular (i.m.) tpa-optimize800 dna vaccine (tpa-s-dna) and boosted with i.p. s-peptide, mice primed with i.m. ctla4hingesars800 dna vaccine (ctla4-s-dn ...	2005	15993989
comparison of 9 different pcr primers for the rapid detection of severe acute respiratory syndrome coronavirus using 2 rna extraction methods.	the sensitivity and specificity of various severe acute respiratory syndrome coronavirus (sars-cov) pcr primer and probe sets were evaluated through the use of commercial kits and in-house amplification formats. conventional and real-time pcr assays were performed using a heat-block thermocycler abi 9600, the roche lightcycler version 1.2, or the abi 7000 sequence detection system. the sensitivity of all primers was between 0.0004 and 0.04 pfu with viral cell lysate and between 0.004 and 0.4 pfu ...	2005	15994050
inhibition of the severe acute respiratory syndrome 3cl protease by peptidomimetic alpha,beta-unsaturated esters.	the proteolytic processing of polyproteins by the 3cl protease of severe acute respiratory syndrome coronavirus is essential for the viral propagation. a series of tripeptide alpha,beta-unsaturated esters and ketomethylene isosteres, including ag7088, are synthesized and assayed to target the 3cl protease. though ag7088 is inactive (ic50 > 100 microm), the ketomethylene isosteres and tripeptide alpha,beta-unsaturated esters containing both p1 and p2 phenylalanine residues show modest inhibitory ...	2005	15994085
sequence analysis and structural prediction of the severe acute respiratory syndrome coronavirus nsp5.	the non-structural proteins (nsp or replicase proteins) of coronaviruses are relatively conserved and can be effective targets for drugs. few studies have been conducted into the function of the severe acute respiratory syndrome coronavirus (sars-cov) nsp5. in this study, bioinformatics methods were employed to predict the secondary structure and construct 3-d models of the sars-cov gd strain nsp5. sequencing and sequential comparison was performed to analyze the mutation trend of the polymerase ...	2005	15999208
differential sensitivities of severe acute respiratory syndrome (sars) coronavirus spike polypeptide enzyme-linked immunosorbent assay (elisa) and sars coronavirus nucleocapsid protein elisa for serodiagnosis of sars coronavirus pneumonia.	the use of recombinant severe acute respiratory syndrome-coronavirus (sars-cov) nucleocapsid protein (n) enzyme-linked immunosorbent assay (elisa)-based antibody and antigen tests for diagnosis of sars-cov infections have been widely reported. however, no recombinant sars-cov spike protein (s)-based elisa is currently available. in this article, we describe the problems and solutions of setting up the recombinant sars-cov s-based elisa for antibody detection. the sars-cov s-based immunoglobulin ...	2005	16000415
evaluation of real-time reverse transcriptase pcr and real-time loop-mediated amplification assays for severe acute respiratory syndrome coronavirus detection.	we compared the performance of a recently established real-time loop-mediated amplification (lamp) assay with the one from a highly sensitive quantitative pcr assay. none of these assays produced false-positive results in this study. for samples isolated from patients within the first 3 days of disease onset, the detection rate of the quantitative pcr assay was higher (14 of 15 were positive) than the lamp assay (9 of 15 were positive). by contrast, the detection rates of these assays toward spe ...	2005	16000477
angiotensin-converting enzyme 2 protects from severe acute lung failure.	acute respiratory distress syndrome (ards), the most severe form of acute lung injury, is a devastating clinical syndrome with a high mortality rate (30-60%) (refs 1-3). predisposing factors for ards are diverse and include sepsis, aspiration, pneumonias and infections with the severe acute respiratory syndrome (sars) coronavirus. at present, there are no effective drugs for improving the clinical outcome of ards. angiotensin-converting enzyme (ace) and ace2 are homologues with different key fun ...	2005	16001071
evaluation of inapparent nosocomial severe acute respiratory syndrome coronavirus infection in vietnam by use of highly specific recombinant truncated nucleocapsid protein-based enzyme-linked immunosorbent assay.	severe acute respiratory syndrome (sars) is a recently emerged human disease associated with pneumonia. inapparent infection with sars coronavirus (cov) is not well characterized. to develop a safe, simple, and reliable screening method for sars diagnosis and epidemiological study, two recombinant sars-cov nucleocapsid proteins (n' protein and (n)delta(121) protein) were expressed in escherichia coli, purified by affinity chromatography, and used as antigens for indirect, immunoglobulin g enzyme ...	2005	16002634
adverse effects of ribavirin and outcome in severe acute respiratory syndrome: experience in two medical centers.	to assess the effect of ribavirin-induced anemia on the outcome of severe acute respiratory syndrome (sars).	2005	16002945
augmentation of immune responses to sars coronavirus by a combination of dna and whole killed virus vaccines.	we studied the immunogenicity of a dna sars-vaccine, a whole killed virus, or a whole killed and dna vaccine combination. the dna vaccine contained a plasmid encoding the sars coronavirus (sars-cov) s protein under the control of the human cmv promoter and intron a. the whole killed virus vaccine was comprised of sars-cov, propagated in vero-e6 cells, with subsequent beta-propilactone inactivation and formulated with aluminum hydroxide adjuvant. mice immunized twice with the dna vaccine and once ...	2005	16005746
a crucial role of angiotensin converting enzyme 2 (ace2) in sars coronavirus-induced lung injury.	during several months of 2003, a newly identified illness termed severe acute respiratory syndrome (sars) spread rapidly through the world. a new coronavirus (sars-cov) was identified as the sars pathogen, which triggered severe pneumonia and acute, often lethal, lung failure. moreover, among infected individuals influenza such as the spanish flu and the emergence of new respiratory disease viruses have caused high lethality resulting from acute lung failure. in cell lines, angiotensin-convertin ...	2005	16007097
protecting human and ecological health under viral threats in asia.	severe acute respiratory syndrome (sars) outbroke in 2003, and the avian influenza a (h5n1) also outbroke in 2003 and continued to 2004. these pandemic viral diseases originated in south east asia. many human and animal lives were lost. economic damages due to the pandemics were also very large. the question arises of why did the pandemics originate from south east asian areas. human influenza a consists of many sub-types of coronaviruses including the sars virus and the avian influenza (h5n1) t ...	2005	16007933
protecting human and ecological health under viral threats in asia.	severe acute respiratory syndrome (sars) outbroke in 2003, and the avian influenza a (h5n1) also outbroke in 2003 and continued to 2004. these pandemic viral diseases originated in south east asia. many human and animal lives were lost. economic damages due to the pandemics were also very large. the question arises of why did the pandemics originate from south east asian areas. human influenza a consists of many sub-types of coronaviruses including the sars virus and the avian influenza (h5n1) t ...	2005	16007933
identification of critical active-site residues in angiotensin-converting enzyme-2 (ace2) by site-directed mutagenesis.	angiotensin-converting enzyme-2 (ace2) may play an important role in cardiorenal disease and it has also been implicated as a cellular receptor for the severe acute respiratory syndrome (sars) virus. the ace2 active-site model and its crystal structure, which was solved recently, highlighted key differences between ace2 and its counterpart angiotensin-converting enzyme (ace), which are responsible for their differing substrate and inhibitor sensitivities. in this study the role of ace2 active-si ...	2005	16008552
the estimation of sars incubation distribution from serial interval data using a convolution likelihood.	the incubation period of sars is the time between infection of disease and onset of symptoms. knowledge about the distribution of incubation times is crucial in determining the length of quarantine period and is an important parameter in modelling the spread and control of sars. as the exact time of infection is unknown for most patients, the incubation time cannot be determined. what is observable is the serial interval which is the time from the onset of symptoms in an index case to the onset ...	2005	16013037
[development of monoclonal antibodies against sars-cov and identification of antigenic epitopes].	based on the genomic sequence of sars-cov strain bj101, antigenic immunodominant genes coding for the structure proteins of sars-cov were predicted by bio-informatics methods, and two chimeric genes a and b with multi-immunodominants lined up by gly-pro-gly linker were synthesized. the chimeric genes were cloned into plasmid pgex-6p-1 and expressed in e. coli with ipgt inducing. balb/c mice were immunized with the purified recombinant fusion protein. the specificity of monoclonal antibodies were ...	2005	16013477
rapid awareness and transmission of severe acute respiratory syndrome in hanoi french hospital, vietnam.	a case-control study was conducted to examine the relationship between severe acute respiratory syndrome (sars) and the time-dependent precautionary behaviors taken during an outbreak of sars in hanoi french hospital (hfh), vietnam. masks (odds ratio [or] = 0.3; 95% confidence interval [ci]: 0.1, 0.7) and gowns (or = 0.2; 95% ci: 0.0, 0.8) appeared to prevent sars transmission. the proportion of doctors and nurses who undertook each measure significantly improved (chi(2) = 9.8551, p = 0.043) aft ...	2005	16014825
apical entry and release of severe acute respiratory syndrome-associated coronavirus in polarized calu-3 lung epithelial cells.	severe acute respiratory syndrome (sars), caused by a novel coronavirus (cov) known as sars-cov, is a contagious and life-threatening respiratory illness with pneumocytes as its main target. a full understanding of how sars-cov would interact with lung epithelial cells will be vital for advancing our knowledge of sars pathogenesis. however, an in vitro model of sars-cov infection using relevant lung epithelial cells is not yet available, making it difficult to dissect the pathogenesis of sars-co ...	2005	16014910
inhibition, escape, and attenuated growth of severe acute respiratory syndrome coronavirus treated with antisense morpholino oligomers.	the recently emerged severe acute respiratory syndrome coronavirus (sars-cov) is a potent pathogen of humans and is capable of rapid global spread. peptide-conjugated antisense morpholino oligomers (p-pmo) were designed to bind by base pairing to specific sequences in the sars-cov (tor2 strain) genome. the p-pmo were tested for their capacity to inhibit production of infectious virus as well as to probe the function of conserved viral rna motifs and secondary structures. several virus-targeted p ...	2005	16014928
the severe acute respiratory syndrome coronavirus 3a protein up-regulates expression of fibrinogen in lung epithelial cells.	here we analyzed the gene expression profile of cells that stably express the severe acute respiratory syndrome coronavirus (sars-cov) 3a protein to determine its effects on host functions. a lung epithelial cell-line, a549, was chosen for this study because the lung is the primary organ infected by sars-cov and fatalities resulted mainly from pulmonary complications. our results showed that the expression of 3a up-regulates the mrna levels of all three subunits, aalpha, bbeta, and gamma, of fib ...	2005	16014971
thrombocytopenia in patients with severe acute respiratory syndrome (review).	severe acute respiratory syndrome (sars) has been recognized as a new human infectious disease caused by a novel coronavirus (sars-cov). hematological changes in patients with sars were common, including notably lymphopenia and thrombocytopenia. while the former is the result of decreases in cd4+ or cd8+ t-lymphocytes related to the onset of disease or use of glucocorticoids, the latter may involve a number of potential mechanisms. although the development of autoimmune antibodies or immune comp ...	2005	16019455
sars vaccine development.	developing effective and safe vaccines is urgently needed to prevent infection by severe acute respiratory syndrome (sars)-associated coronavirus (sars-cov). the inactivated sars-cov vaccine may be the first one available for clinical use because it is easy to generate; however, safety is the main concern. the spike (s) protein of sars-cov is the major inducer of neutralizing antibodies, and the receptor-binding domain (rbd) in the s1 subunit of s protein contains multiple conformational neutral ...	2005	16022774
sars coronavirus detection methods.	using clinical samples from patients with severe acute respiratory syndrome, we showed that the sensitivities of a quantitative reverse transcription-polymerase chain reaction (80% for fecal samples and 25% for urine samples) were higher than those of the polyclonal (50% and 5%) and monoclonal (35% and 8%) antibody-based nucleocapsid antigen capture enzyme-linked immunosorbent assays.	2005	16022791
asymptomatic sars coronavirus infection among healthcare workers, singapore.	we conducted a study among healthcare workers (hcws) exposed to patients with severe acute respiratory syndrome (sars) before infection control measures were instituted. of all exposed hcws, 7.5% had asymptomatic sars-positive cases. asymptomatic sars was associated with lower sars antibody titers and higher use of masks when compared to pneumonic sars.	2005	16022801
adenoviral expression of a truncated s1 subunit of sars-cov spike protein results in specific humoral immune responses against sars-cov in rats.	the causative agent of severe acute respiratory syndrome (sars) has been identified as sars-associated coronavirus (sars-cov), but the prophylactic treatment of sars-cov is still under investigation. we constructed a recombinant adenovirus containing a truncated n-terminal fragment of the sars-cov spike (s) gene (from--45 to 1469, designated ad-s(n)), which encoded a truncated s protein (490 amino-acid residues, a part of 672 amino-acid s1 subunit), and investigated whether this construct could ...	2005	16022898
thermal aggregation of sars-cov membrane protein.	sars-cov membrane protein could be detected easily using western blotting in non-denaturing condition but not regular denaturing treatment. boiling treatment, causing the aggregation of sars-cov membrane protein in the stacking gels, results in the failure to detect the membrane protein in the separating gels. aggregated membrane proteins could not be dissociated by 1% triton-x 100, 6m urea, or 2% sds. the region with amino acid residues from 51 to 170 is responsible for thermal aggregation of s ...	2005	16023741
characterization of viral proteins encoded by the sars-coronavirus genome.	a new disease, termed severe acute respiratory syndrome (sars), emerged at the end of 2002 and caused profound disturbances in over 30 countries worldwide in 2003. a novel coronavirus was identified as the aetiological agent of sars and the 30kb viral genome was deciphered with unprecedented speed in a coordinated manner by the global community. since then, much progress has been made in the virological and molecular characterization of the proteins encoded by sars-coronavirus (sars-cov) genome, ...	2005	15708633
recombinant modified vaccinia virus ankara expressing the spike glycoprotein of severe acute respiratory syndrome coronavirus induces protective neutralizing antibodies primarily targeting the receptor binding region.	immunization with a killed or inactivated viral vaccine provides significant protection in animals against challenge with certain corresponding pathogenic coronaviruses (covs). however, the promise of this approach in humans is hampered by serious concerns over the risk of leaking live severe acute respiratory syndrome (sars) viruses. in this study, we generated a sars vaccine candidate by using the live-attenuated modified vaccinia virus ankara (mva) as a vector. the full-length sars-cov envelo ...	2005	15708987
recombinant modified vaccinia virus ankara expressing the spike glycoprotein of severe acute respiratory syndrome coronavirus induces protective neutralizing antibodies primarily targeting the receptor binding region.	immunization with a killed or inactivated viral vaccine provides significant protection in animals against challenge with certain corresponding pathogenic coronaviruses (covs). however, the promise of this approach in humans is hampered by serious concerns over the risk of leaking live severe acute respiratory syndrome (sars) viruses. in this study, we generated a sars vaccine candidate by using the live-attenuated modified vaccinia virus ankara (mva) as a vector. the full-length sars-cov envelo ...	2005	15708987