Publications

generation and characterization of dna vaccines targeting the nucleocapsid protein of severe acute respiratory syndrome coronavirus.	severe acute respiratory syndrome (sars) is a serious threat to public health and the economy on a global scale. the sars coronavirus (sars-cov) has been identified as the etiological agent for sars. thus, vaccination against sars-cov may represent an effective approach to controlling sars. dna vaccines are an attractive approach for sars vaccine development, as they offer many advantages over conventional vaccines, including stability, simplicity, and safety. our investigators have previously s ...	2004	15078946
evaluation of advanced reverse transcription-pcr assays and an alternative pcr target region for detection of severe acute respiratory syndrome-associated coronavirus.	first-generation reverse transcription-pcr (rt-pcr) assays for severe acute respiratory syndrome-associated coronavirus (sars-cov) gave false-negative results in a considerable fraction of patients. in the present study, we evaluated two second-generation, replicase (r) gene-based, real-time rt-pcr test kits--the realart hpa coronavirus lc kit (artus, hamburg, germany) and the lightcycler sars-cov quantification kit (roche, penzberg, germany)--and a real-time rt-pcr assay for the nucleocapsid (n ...	2004	15131168
the involvement of natural killer cells in the pathogenesis of severe acute respiratory syndrome.	by using peripheral blood samples from 221 cases of severe acute respiratory syndrome (sars), 34 of mycoplasma pneumoniae infection, and 44 healthy adults, we measured the total number of natural killer (nk) and cd158b+ nk cells (cd158b+ nk) using flow cytometric analysis and calculated the percentage of cd158b+ nk cells. the total number of nk and cd158b+ nk cells and the percentage of cd158b+ nk cells were significantly lower in patients with sars than in those with m. pneumoniae infection (p ...	2004	15080302
detection of severe acute respiratory syndrome-associated coronavirus in pneumocytes of the lung.	previous reports have indicated that patients with severe acute respiratory syndrome (sars)-associated coronavirus infection could develop atypical pneumonia with fulminant pulmonary edema. however, the target cells of sars viral infection have not been characterized in detail. we report the pathologic findings of the lung in 3 cases of sars. chest radiographs at 2 to 3 weeks of infection revealed an atypical pneumonia with pulmonary consolidation, a clinical characteristic of sars infection. th ...	2004	15080310
induction of sars-nucleoprotein-specific immune response by use of dna vaccine.	induction of effective cytotoxic t lymphocyte (ctl) and/or a specific antibody against conserved viral proteins may be essential to the development of a safe and effective severe acute respiratory syndrome coronavirus (sars-cov) vaccine. dna vaccination represents a new strategy for induction of humoral and cellular immune response. to determine the ability of sars-cov nucleoprotein (n protein) to induce antiviral immunity, in this report, we established a stable c2c12 line expressing sars-cov n ...	2004	15081618
are there characteristics of infectious diseases that raise special ethical issues?	this paper examines the characteristics of infectious diseases that raise special medical and social ethical issues, and explores ways of integrating both current bioethical and classical public health ethics concerns. many of the ethical issues raised by infectious diseases are related to these diseases' powerful ability to engender fear in individuals and panic in populations. we address the association of some infectious diseases with high morbidity and mortality rates, the sense that infecti ...	2004	15086371
primary characterization of sars coronavirus strain frankfurt 1.		2004	15088406
the nucleocapsid protein of the sars coronavirus is capable of self-association through a c-terminal 209 amino acid interaction domain.	severe acute respiratory syndrome (sars) coronavirus (sars-cov) caused a severe outbreak in several regions of the world in 2003. the virus is a novel coronavirus isolated from patients exhibiting atypical pneumonia and may have originated from wild animals such as civet cats in southern china. the genome of sars-cov is a positive-sense, single-stranded rna whose sequence is distantly related to all known coronaviruses that infect humans and animals. like other known coronaviruses, sars-cov is a ...	2004	15094372
[sars].		2004	15095764
antibody response and viraemia during the course of severe acute respiratory syndrome (sars)-associated coronavirus infection.	to understand the time-course of viraemia and antibody responses to severe acute respiratory syndrome-associated coronavirus (sars-cov), rt-pcr and elisa were used to assay 376 blood samples from 135 sars patients at various stages of the illness, including samples from patients who were in their early convalescent phase. the results showed that igm antibodies decreased and became undetectable 11 weeks into the recovery phase. igg antibodies, however, remained detectable for a period beyond 11 w ...	2004	15096554
the antiviral effect of interferon-beta against sars-coronavirus is not mediated by mxa protein.	severe acute respiratory syndrome (sars) is caused by a novel coronavirus termed sars-cov. no antiviral treatment has been established so far. interferons are cytokines which induce the synthesis of several antivirally active proteins in the cell. in this study, we demonstrated that multiplication of sars-cov in cell culture can be strongly inhibited by pretreatment with interferon-beta. interferon-alpha and interferon-gamma, by contrast, were less effective. the human mxa protein is one of the ...	2004	15135736
a one step quantitative rt-pcr for detection of sars coronavirus with an internal control for pcr inhibitors.	in this study, we report a one step quantitative rt-pcr assay for severe acute respiratory syndrome (sars) diagnosis. the overall detection rate for clinical samples collected from days 1 to 9 of disease onset is 86.2% and the specificity of the assay is 100%. to detect false negative results due to pcr inhibitors or faulty rna extraction, we have further modified this one step rt-pcr assay for simultaneous detection of severe acute respiratory syndrome (sars) coronavirus (cov) and 18s ribosomal ...	2004	15135737
severe acute respiratory syndrome coronavirus spike protein expressed by attenuated vaccinia virus protectively immunizes mice.	the spike protein (s), a membrane component of severe acute respiratory syndrome coronavirus (sars-cov) is anticipated to be an important component of candidate vaccines. we constructed recombinant forms of the highly attenuated modified vaccinia virus ankara (mva) containing the gene encoding full-length sars-cov s with and without a c-terminal epitope tag called mva/s-ha and mva/s, respectively. cells infected with mva/sor mva/s-ha synthesized a 200-kda protein, which was recognized by antibod ...	2004	15096611
evidence of airborne transmission of the severe acute respiratory syndrome virus.	there is uncertainty about the mode of transmission of the severe acute respiratory syndrome (sars) virus. we analyzed the temporal and spatial distributions of cases in a large community outbreak of sars in hong kong and examined the correlation of these data with the three-dimensional spread of a virus-laden aerosol plume that was modeled using studies of airflow dynamics.	2004	15102999
multiple enzymatic activities associated with severe acute respiratory syndrome coronavirus helicase.	severe acute respiratory syndrome coronavirus (sars-cov), a newly identified group 2 coronavirus, is the causative agent of severe acute respiratory syndrome, a life-threatening form of pneumonia in humans. coronavirus replication and transcription are highly specialized processes of cytoplasmic rna synthesis that localize to virus-induced membrane structures and were recently proposed to involve a complex enzymatic machinery that, besides rna-dependent rna polymerase, helicase, and protease act ...	2004	15140959
laboratory-acquired severe acute respiratory syndrome.		2004	15103000
sars ctl vaccine candidates; hla supertype-, genome-wide scanning and biochemical validation.	an effective severe acute respiratory syndrome (sars) vaccine is likely to include components that can induce specific cytotoxic t-lymphocyte (ctl) responses. the specificities of such responses are governed by human leukocyte antigen (hla)-restricted presentation of sars-derived peptide epitopes. exact knowledge of how the immune system handles protein antigens would allow for the identification of such linear sequences directly from genomic/proteomic sequence information (lauemoller et al., re ...	2004	15104671
identification of novel inhibitors of the sars coronavirus main protease 3clpro.	sars (severe acute respiratory syndrome) is caused by a newly discovered coronavirus. a key enzyme for the maturation of this virus and, therefore, a target for drug development is the main protease 3cl(pro) (also termed sars-cov 3cl(pro)). we have cloned and expressed in escherichia coli the full-length sars-cov 3cl(pro) as well as a truncated form containing only the catalytic domains. the recombinant proteins have been characterized enzymatically using a fluorescently labeled substrate; their ...	2004	15109248
coronaviridae and sars-associated coronavirus strain hsr1.	during the recent severe acute respiratory (sars) outbreak, the etiologic agent was identified as a new coronavirus (cov). we have isolated a sars-associated cov (sars-cov) strain by injecting vero cells with a sputum specimen from an italian patient affected by a severe pneumonia; the patient traveled from vietnam to italy in march 2003. ultrastructural analysis of infected vero cells showed the virions within cell vesicles and around the cell membrane. the full-length viral genome sequence was ...	2004	15109406
development of taqman rt-nested pcr system for clinical sars-cov detection.	severe acute respiratory syndrome (sars) is an acute newly emerged infectious respiratory illness. the etiologic agent of sars was named 'sars-associated coronavirus' (sars-cov) that can be detected with reverse transcription-polymerase chain reaction (rt-pcr) assays. in this study, 12 sets of nested primers covering the sars-cov genome have been screened and showed sufficient sensitivity to detect sars-cov in rna isolated from virus cultured in vero 6 cells. to optimize further the reaction con ...	2004	15109816
microbicides and the environmental control of nosocomial viral infections.	viruses are important causes of acute and chronic diseases in humans. newer viruses are still being discovered and those that are already known are being incriminated in the aetiology of clinical conditions with hitherto unknown causes. apart from frequently causing infections in the general community, many types of viruses are also significant nosocomial pathogens. while it is generally agreed that we underestimate the proportion of nosocomial infections that are viral, due to a lack of routine ...	2004	15110126
dissection study on the severe acute respiratory syndrome 3c-like protease reveals the critical role of the extra domain in dimerization of the enzyme: defining the extra domain as a new target for design of highly specific protease inhibitors.	the severe acute respiratory syndrome (sars) 3c-like protease consists of two distinct folds, namely the n-terminal chymotrypsin fold containing the domains i and ii hosting the complete catalytic machinery and the c-terminal extra helical domain iii unique for the coronavirus 3cl proteases. previously the functional role of this extra domain has been completely unknown, and it was believed that the coronavirus 3cl proteases share the same enzymatic mechanism with picornavirus 3c proteases, whic ...	2004	15037623
analysis of synonymous codon usage in sars coronavirus and other viruses in the nidovirales.	in this study, we calculated the codon usage bias in severe acute respiratory syndrome coronavirus (sarscov) and performed a comparative analysis of synonymous codon usage patterns in sarscov and 10 other evolutionary related viruses in the nidovirales. although there is a significant variation in codon usage bias among different sarscov genes, codon usage bias in sarscov is a little slight, which is mainly determined by the base compositions on the third codon position. by comparing synonymous ...	2004	15041183
exploring the binding mechanism of the main proteinase in sars-associated coronavirus and its implication to anti-sars drug design.	the main proteinase of sars-associated coronavirus (sars-cov) plays an important role in viral transcription and replication, and is an attractive target for anti-sars drug development. the important thing is to understand its binding mechanism with possible ligands. in this study, we investigated possible noncanonical interactions, potential inhibitors, and binding pockets in the main proteinase of sars-cov based on its recently determined crystal structure. these findings provide a wide clue t ...	2004	15080921
[epidemiological investigation of nosocomial infection of sars in medical staff of a hospital].	to investigate the epidemiological features of nosocomial infection of severe acute respiratory syndrome (sars) in the medical staff of a hospital.	2004	15041564
[severe acute respiratory syndrome: a singular epidemic of viral pneumonia].	infectious agent: the severe acute respiratory syndrome (sars) is a febrile pneumonia initially observed in china at the end of 2002. the infectious agent has rapidly been identified as a new coronavirus, baptised sars-associated coronavirus (cov-sars). transmission is inter-human, via respiratory particles mainly. clinical presentation and treatment: the clinical presentation is highly variable, from a mild fever to an acute respiratory distress syndrome. there is no specific treatment. ribavir ...	2004	15041887
sars virus returns to china as scientists race to find effective vaccine.		2004	15042239
severe acute respiratory syndrome: scientific and anecdotal evidence for drug treatment.	severe acute respiratory syndrome (sars), caused by a highly infectious novel coronavirus (cov), predominantly presents with severe pneumonitis leading to respiratory failure and death in approximately 10% of victims. most cases present, after an incubation of 2 to 11 days, with fever and chills, which are followed by dry cough and dyspnea before the onset of respiratory failure. the management of sars is controversial, largely due to the lack of data from controlled trials, which were logistica ...	2004	15043392
viral attacks on the blood supply: the impact of severe acute respiratory syndrome in beijing.		2004	15043559
severe acute respiratory syndrome.	severe acute respiratory syndrome (sars) is a new respiratory illness due to a novel coronavirus called sars-cov. cases of sars were first identified from guangdong province in southern china in november 2002. over the next several months, international travel allowed for the rapid spread of >8,000 cases over three continents. the economic and psychological impact of this mysterious illness was profound. in order to better educate the public about this emerging infectious disease, we describe th ...	2004	15095827
nat screening of blood donors for severe acute respiratory syndrome coronavirus can potentially prevent transfusion associated transmissions.	the severe acute respiratory syndrome (sars) was first described in february 2003. close contact with symptomatic patients appears to be the main route of transmission, whereas blood transfusion transmission could not be ruled out.	2004	15043560
coronavirus outbreak in cheetahs: lessons for sars.		2004	15043830
sars-associated coronavirus quasispecies in individual patients.		2004	15044654
mutational patterns correlate with genome organization in sars and other coronaviruses.	focused efforts by several international laboratories have resulted in the sequencing of the genome of the causative agent of severe acute respiratory syndrome (sars), novel coronavirus sars-cov, in record time. using cumulative skew diagrams, i found tht mutational patterns in the sars-cov genome were strikingly different from other coronaviruses in terms of mutation rates, although they were in general agreement with the model of the coronavirus lifecycle. these findings might be relevant for ...	2004	15049309
rapid detection of the severe acute respiratory syndrome (sars) coronavirus by a loop-mediated isothermal amplification assay.		2004	15054079
antigenicity analysis of different regions of the severe acute respiratory syndrome coronavirus nucleocapsid protein.	the widespread threat of severe acute respiratory syndrome (sars) to human health has made urgent the development of fast and accurate analytical methods for its early diagnosis and a safe and efficient antiviral vaccine for preventive use. for this purpose, we investigated the antigenicity of different regions of the sars coronavirus (sars-cov) nucleocapsid (n) protein.	2004	15054081
bioterrorism and emerging infectious disease - antimicrobials, therapeutics and immune-modulators. sars coronavirus.	the purpose of this meeting was to provide a forum for expert presentations and discussion about the threats of bioterrorism and emerging infectious diseases, and to address the issues relating to epidemics, prevention of infection and treatment of some of these emerging infectious diseases classified as potential agents of bioterror. included in the talks were state-of-the-art presentations about infectious clone technology and recombinant viruses, pathogen and receptor interactions at the cell ...	2004	15057645
[severe acute respiratory syndrome].		2004	15108450
[an epidemiologic investigation on infection with severe acute respiratory syndrome coronavirus in wild animals traders in guangzhou].	to investigate status of infection with severe acute respiratory syndrome coronovirus (sars-cov) in traders of wild animals wholesale markets in guangzhou.	2004	15061910
[detection for severe acute respiratory syndrome (sars) coronavirus rna in stool of sars patients].	to investigate excretion of severe acute respiratory syndrome coronavirus rna (sars-cov) in stool of sars patients.	2004	15061913
performance and cost evaluation of one commercial and six in-house conventional and real-time reverse transcription-pcr assays for detection of severe acute respiratory syndrome coronavirus.	we evaluated seven reverse transcription-pcr (rt-pcr) assays, including six in-house assays and one commercial assay for the detection of severe acute respiratory syndrome coronavirus (sars-cov) rna in clinical specimens. rt-pcr assays targeted different genomic regions and included three conventional assays (one nested and two non-nested) run on a conventional heat block and four real-time assays performed in a lightcycler (lc; roche diagnostics). all in-house assays were optimized for assay pa ...	2004	15070991
factors that make an infectious disease outbreak controllable.	the aim of this study is to identify general properties of emerging infectious agents that determine the likely success of two simple public health measures in controlling outbreaks, namely (i) isolating symptomatic individuals and (ii) tracing and quarantining their contacts. because these measures depend on the recognition of specific disease symptoms, we investigate the relative timing of infectiousness and the appearance of symptoms by using a mathematical model. we show that the success of ...	2004	15071187
old drugs as lead compounds for a new disease? binding analysis of sars coronavirus main proteinase with hiv, psychotic and parasite drugs.	the sars-associated coronavirus (sars-cov) main proteinase is a key enzyme in viral polyprotein processing. to allow structure-based design of drugs directed at sars-cov main proteinase, we predicted its binding pockets and affinities with existing hiv, psychotic and parasite drugs (lopinavir, ritonavir, niclosamide and promazine), which show signs of inhibiting the replication of sars-cov. our results suggest that these drugs and another two hiv inhibitors (pnu and uc2) could be used as templat ...	2004	15110833
severe acute respiratory syndrome: report of treatment and outcome after a major outbreak.	the outcome is reported of a prospective uncontrolled study based on a stepwise treatment protocol during an outbreak of severe acute respiratory syndrome (sars) in hong kong.	2004	15115870
monophyletic relationship between severe acute respiratory syndrome coronavirus and group 2 coronaviruses.	although primary genomic analysis has revealed that severe acute respiratory syndrome coronavirus (sars cov) is a new type of coronavirus, the different protein trees published in previous reports have provided no conclusive evidence indicating the phylogenetic position of sars cov. to clarify the phylogenetic relationship between sars cov and other coronaviruses, we compiled a large data set composed of 7 concatenated protein sequences and performed comprehensive analyses, using the maximum-lik ...	2004	15116304
evidence from the evolutionary analysis of nucleotide sequences for a recombinant history of sars-cov.	the origins and evolutionary history of the severe acute respiratory syndrome (sars) coronavirus (sars-cov) remain an issue of uncertainty and debate. based on evolutionary analyses of coronavirus dna sequences, encompassing an approximately 13kb stretch of the sars-tor2 genome, we provide evidence that sars-cov has a recombinant history with lineages of types i and iii coronavirus. we identified a minimum of five recombinant regions ranging from 83 to 863bp in length and including the polymeras ...	2004	15019585
analysis of multimerization of the sars coronavirus nucleocapsid protein.	severe acute respiratory syndrome (sars), an emerging disease characterized by atypical pneumonia, has recently been attributed to a novel coronavirus. the genome of sars coronavirus (sars-cov) has recently been sequenced, and a number of genes identified, including that of the nucleocapsid protein (n). it is noted, however, that the n protein of sars-cov (sars-cov n) shares little homology with nucleocapsid proteins of other members of the coronavirus family [science 300 (2003) 1399; science 30 ...	2004	15020242
a dna vaccine induces sars coronavirus neutralization and protective immunity in mice.	public health measures have successfully identified and contained outbreaks of the severe acute respiratory syndrome (sars) coronavirus (sars-cov), but concerns remain over the possibility of future recurrences. finding a vaccine for this virus therefore remains a high priority. here, we show that a dna vaccine encoding the spike (s) glycoprotein of the sars-cov induces t cell and neutralizing antibody responses, as well as protective immunity, in a mouse model. alternative forms of s were analy ...	2004	15024391
radiologic pattern of disease in patients with severe acute respiratory syndrome: the toronto experience.	severe acute respiratory syndrome (sars) is a transmissible febrile respiratory illness caused by a recently discovered coronavirus. various patterns of disease progression may be observed that have different implications for the prognosis in those affected by sars. the appearance of the lungs on chest radiographs of patients with this condition may be normal or may include focal airspace opacity or multifocal or diffuse opacities. thoracic computed tomography (ct) is more sensitive in depicting ...	2004	15026600
sars-related virus predating sars outbreak, hong kong.	using immunofluorescence and neutralization assays, we detected antibodies to human severe acute respiratory syndrome-associated coronavirus (sars-cov) and/or animal sars-cov-like virus in 17 (1.8%) of 938 adults recruited in 2001. this finding suggests that a small proportion of healthy persons in hong kong had been exposed to sars-related viruses at least 2 years before the recent sars outbreak.	2004	15030679
susceptibility of pigs and chickens to sars coronavirus.	an outbreak of severe acute respiratory syndrome (sars) in humans, associated with a new coronavirus, was reported in southeast asia, europe, and north america in early 2003. to address speculations that the virus originated in domesticated animals, or that domestic species were susceptible to the virus, we inoculated 6-week-old pigs and chickens intravenously, intranasally, ocularly, and orally with 106 pfu of sars-associated coronavirus (sars-cov). clinical signs did not develop in any animal, ...	2004	15030680
susceptibility of pigs and chickens to sars coronavirus.	an outbreak of severe acute respiratory syndrome (sars) in humans, associated with a new coronavirus, was reported in southeast asia, europe, and north america in early 2003. to address speculations that the virus originated in domesticated animals, or that domestic species were susceptible to the virus, we inoculated 6-week-old pigs and chickens intravenously, intranasally, ocularly, and orally with 106 pfu of sars-associated coronavirus (sars-cov). clinical signs did not develop in any animal, ...	2004	15030680
sars surveillance during emergency public health response, united states, march-july 2003.	in response to the emergence of severe acute respiratory syndrome (sars), the united states established national surveillance using a sensitive case definition incorporating clinical, epidemiologic, and laboratory criteria. of 1,460 unexplained respiratory illnesses reported by state and local health departments to the centers for disease control and prevention from march 17 to july 30, 2003, a total of 398 (27%) met clinical and epidemiologic sars case criteria. of these, 72 (18%) were probable ...	2004	15030681
introduction of sars in france, march-april, 2003.	we describe severe acute respiratory syndrome (sars) in france. patients meeting the world health organization definition of a suspected case underwent a clinical, radiologic, and biologic assessment at the closest university-affiliated infectious disease ward. suspected cases were immediately reported to the institut de veille sanitaire. probable case-patients were isolated, their contacts quarantined at home, and were followed for 10 days after exposure. five probable cases occurred from march ...	2004	15030682
sars-associated coronavirus transmission, united states.	to better assess the risk for transmission of the severe acute respiratory syndrome-associated coronavirus (sars-cov), we obtained serial specimens and clinical and exposure data from seven confirmed u.s. sars patients and their 10 household contacts. sars-cov was detected in a day-14 sputum specimen from one case-patient and in five stool specimens from two case-patients. in one case-patient, sars-cov persisted in stool for at least 26 days after symptom onset. the highest amounts of virus were ...	2004	15030687
healthcare worker seroconversion in sars outbreak.	serum samples were obtained from healthcare workers 5 weeks after exposure to an outbreak of severe acute respiratory syndrome (sars). a sensitive dot blot enzyme-linked immunosorbent assay, complemented by a specific neutralization test, shows that only persons in whom probable sars was diagnosed had specific antibodies and suggests that subclinical sars is not an important feature of the disease.	2004	15030691
surgical helmets and sars infection.	performance testing of two brands of surgical helmets indicated that their efficiency at in vivo filtration of sub-micrometer-sized particles is inadequate for their use as respirators. these helmets are not marketed for respiratory protection and should not be used alone for protection against severe acute respiratory syndrome when performing aerosol-generating procedures.	2004	15030697
possible sars coronavirus transmission during cardiopulmonary resuscitation.	infection of healthcare workers with the severe acute respiratory syndrome-associated coronavirus (sars-cov) is thought to occur primarily by either contact or large respiratory droplet transmission. however, infrequent healthcare worker infections occurred despite the use of contact and droplet precautions, particularly during certain aerosol-generating medical procedures. we investigated a possible cluster of sars-cov infections in healthcare workers who used contact and droplet precautions du ...	2004	15030699
detection of sars coronavirus in patients with suspected sars.	cases of severe acute respiratory syndrome (sars) were investigated for sars coronavirus (sars-cov) through rna tests, serologic response, and viral culture. of 537 specimens from patients in whom sars was clinically diagnosed, 332 (60%) had sars-cov rna in one or more clinical specimens, compared with 1 (0.3%) of 332 samples from controls. of 417 patients with clinical sars from whom paired serum samples were available, 92% had an antibody response. rates of viral rna positivity increased progr ...	2004	15030700
interaction between heptad repeat 1 and 2 regions in spike protein of sars-associated coronavirus: implications for virus fusogenic mechanism and identification of fusion inhibitors.	studies on the fusion-inhibitory peptides derived from the heptad repeat 1 and 2 (hr1 and hr2) regions of the hiv-1 envelope glycoprotein gp41 provided crucial information on the viral fusogenic mechanism. we used a similar approach to study the fusogenic mechanism of severe-acute-respiratory-syndrome-associated coronavirus (sars-cov).	2004	15043961
synthetic peptide studies on the severe acute respiratory syndrome (sars) coronavirus spike glycoprotein: perspective for sars vaccine development.	the s (spike) protein of the etiologic coronavirus (cov) agent of severe acute respiratory syndrome (sars) plays a central role in mediating viral infection via receptor binding and membrane fusion between the virion and the host cell. we focused on using synthetic peptides for developing antibodies against sars-cov, which aimed to block viral invasion by eliciting an immune response specific to the native sars-cov s protein.	2004	15044316
real-time polymerase chain reaction for detecting sars coronavirus, beijing, 2003.	during the 2003 severe acute respiratory syndrome (sars) outbreak, a real-time quantitative polymerase chain reaction, which targets the nucleocapsid gene at the 3' end of the viral genome, was established to detect and identify the sars-associated coronavirus. we describe the use of this assay to screen >700 clinical samples.	2004	15030701
serologic and molecular biologic methods for sars-associated coronavirus infection, taiwan.	severe acute respiratory syndrome (sars) has raised a global alert since march 2003. after its causative agent, sars-associated coronavirus (sars-cov), was confirmed, laboratory methods, including virus isolation, reverse transcriptase-polymerase chain reaction (rt-pcr), and serologic methods, have been quickly developed. in this study, we evaluated four serologic tests ( neutralization test, enzyme-linked immunosorbent assay [elisa], immunofluorescent assay [ifa], and immunochromatographic test ...	2004	15030702
one-tube nested rt-pcr enabled by using a plastic film and its application for the rapid detection of sars-virus.	a general strategy based on the use of a plastic film that allows reverse transcription and nested-pcr in a single closed-tube has been developed. the reaction mixture for the second pcr amplification is quarantined in the cap of the reaction tube during the first round amplification by a piece of plastic film, and later introduced into the pcr amplicons from the first round reaction by centrifugation without opening the reaction tube. the main advantages of our method are its high sensitivity, ...	2004	15049359
real-time reverse transcription-polymerase chain reaction assay for sars-associated coronavirus.	a real-time reverse transcription-polymerase chain reaction (rt-pcr) assay was developed to rapidly detect the severe acute respiratory syndrome-associated coronavirus (sars-cov). the assay, based on multiple primer and probe sets located in different regions of the sars-cov genome, could discriminate sars-cov from other human and animal coronaviruses with a potential detection limit of <10 genomic copies per reaction. the real-time rt-pcr assay was more sensitive than a conventional rt-pcr assa ...	2004	15030703
interferon-beta 1a and sars coronavirus replication.	a global outbreak of severe acute respiratory syndrome (sars) caused by a novel coronavirus began in march 2003. the rapid emergence of sars and the substantial illness and death it caused have made it a critical public health issue. because no effective treatments are available, an intensive effort is under way to identify and test promising antiviral drugs. here, we report that recombinant human interferon-beta 1a potently inhibits sars coronavirus replication in vitro.	2004	15030704
ultrastructural characterization of sars coronavirus.	severe acute respiratory syndrome (sars) was first described during a 2002-2003 global outbreak of severe pneumonia associated with human deaths and person-to-person disease transmission. the etiologic agent was initially identified as a coronavirus by thin-section electron microscopic examination of a virus isolate. virions were spherical, 78 nm in mean diameter, and composed of a helical nucleocapsid within an envelope with surface projections. we show that infection with the sars-associated c ...	2004	15030705
phylogenomics and bioinformatics of sars-cov.		2004	15058277
combining clinical and epidemiologic features for early recognition of sars.	early recognition and rapid initiation of infection control precautions are currently the most important strategies for controlling severe acute respiratory syndrome (sars). no rapid diagnostic tests currently exist that can rule out sars among patients with febrile respiratory illnesses. clinical features alone cannot with certainty distinguish sars from other respiratory illnesses rapidly enough to inform early management decisions. a balanced approach to screening that allows early recognitio ...	2004	15030706
possible central nervous system infection by sars coronavirus.	on day 22 of illness, generalized tonic-clonic convulsion developed in a 32-year-old woman with severe acute respiratory syndrome (sars). cerebrospinal fluid tested positive for sars coronavirus (sars-cov) by reverse transcriptase-polymerase chain reaction. sars-cov may have caused an infection in the central nervous system in this patient.	2004	15030709
procalcitonin in severe acute respiratory syndrome (sars).	the role of procalcitonin (pct) in severe acute respiratory syndrome (sars) has not been highlighted so far. we described retrospectively eight cases of sepsis from pneumonia of various microbiological aetiologies including two due to sars, compared their pct concentrations and provided further descriptors of sars as a viral pneumonia.	2004	15066330
sars and pregnancy: a case report.	we report a laboratory-confirmed case of severe acute respiratory syndrome (sars) in a pregnant woman. although the patient had respiratory failure, a healthy infant was subsequently delivered, and the mother is now well. there was no evidence of viral shedding at delivery. antibodies to sars virus were detected in cord blood and breast milk.	2004	15030710
immunological characterization of the spike protein of the severe acute respiratory syndrome coronavirus.	severe acute respiratory syndrome (sars) is a novel infectious disease caused by the sars-associated coronavirus (sars-cov). there are four major structural proteins in the sars-cov, including the nucleocapsid, spike, membrane, and small envelope proteins. in this study, two sets of truncated fragments of spike protein were generated, the first were approximately 210-bp nonoverlapping fragments and the second were overlapping segments of 750 to 900 bp. from these 23 fragments, we identified a fr ...	2004	15071006
boosting the sensitivity of real-time polymerase-chain-reaction testing for sars.		2004	15071137
atypical sars and escherichia coli bacteremia.	we describe a patient with severe acute respiratory syndrome (sars) whose clinical symptoms were masked by escherichia coli bacteremia. sars developed in a cluster of healthcare workers who had contact with this patient. sars was diagnosed when a chest infiltrate developed and when the patient's brother was hospitalized with acute respiratory failure. we highlight problems in atypical cases and offer infection control suggestions.	2004	15030711
relative rates of non-pneumonic sars coronavirus infection and sars coronavirus pneumonia.	although the genome of severe acute respiratory syndrome coronavirus (sars-cov) has been sequenced and a possible animal reservoir identified, seroprevalence studies and mass screening for detection of subclinical and non-pneumonic infections are still lacking.	2004	15031027
serology of severe acute respiratory syndrome: implications for surveillance and outcome.	severe acute respiratory syndrome (sars) is a novel infectious disease. no information is currently available on host-specific immunity against the sars coronavirus (cov), and detailed characteristics of the epidemiology of sars cov infection have not been identified.	2004	15031782
inhibition of severe acute respiratory syndrome-associated coronavirus (sarscov) by calpain inhibitors and beta-d-n4-hydroxycytidine.	we evaluated two types of compounds for efficacy in inhibiting sarscov replication in vitro: calpain inhibitors (a class of cellular cysteine proteinases) and a number of nucleoside analogues. cytopathic effect reduction assays visually determined with spectrophotometric verification by neutral red (nr) uptake assay were used to evaluate cytotoxicity and antiviral potency of the compounds. significantly inhibitory compounds were then evaluated in virus yield reduction assays. two calpain inhibit ...	2004	15074711
severe acute respiratory syndrome-related coronavirus is inhibited by interferon- alpha.	current treatment schemes for severe acute respiratory syndrome (sars) include broad-spectrum antibiotics, glucocorticoids, and ribavirin. we evaluated the susceptibility of the sars-related coronavirus (sars cov) to ribavirin and interferon (ifn)- alpha in vitro by use of cytopathic effect, plaque assay, and immunoblot analysis. ribavirin did not inhibit viral growth at concentrations attainable in human serum. in contrast, ifn- alpha showed an in vitro inhibitory effect starting at concentrati ...	2004	15031783
severe acute respiratory syndrome-related coronavirus is inhibited by interferon- alpha.	current treatment schemes for severe acute respiratory syndrome (sars) include broad-spectrum antibiotics, glucocorticoids, and ribavirin. we evaluated the susceptibility of the sars-related coronavirus (sars cov) to ribavirin and interferon (ifn)- alpha in vitro by use of cytopathic effect, plaque assay, and immunoblot analysis. ribavirin did not inhibit viral growth at concentrations attainable in human serum. in contrast, ifn- alpha showed an in vitro inhibitory effect starting at concentrati ...	2004	15031783
protein structure prediction in structure based drug design.	the human genome and other genome sequencing projects have generated huge amounts of protein sequence information. recently, a structural genomics project that aims to determine at least one representative three-dimensional structure from every protein family experimentally has been started. homology modeling will play an essential role in structure based drug design such as in silico screening; because based on these representative structures the three-dimensional structures of the remaining pr ...	2004	15032603
[comparative study of clinical characteristics and prognosis of clinically diagnosed sars patients with positive and negative serum sars coronavirus-specific antibodies test].	to investigate clinical characteristics and long-term effects of sars.	2004	15130303
[new developments as regards coronaviruses].		2004	15134085
probing the structure of the sars coronavirus using scanning electron microscopy.	a novel coronavirus, sars-cov, has been confirmed to be the aetiological agent of sars. transmission electron microscope (tem) images played an important role in initial identification of the pathogen. in order to obtain greater morphological detail of sars-cov than could be obtained by tem, we used ultra-high resolution scanning electron microscopy (sem) to image the virus particles. we show here the three-dimensional appearance of sars-cov. enhanced detail of the ultrastructure reveals the tri ...	2004	15134191
identification of a novel protein 3a from severe acute respiratory syndrome coronavirus.	the open reading frame 3 of the severe acute respiratory syndrome coronavirus (sars-cov) genome encodes a predicted protein 3a, consisting of 274 amino acids, that lacks any significant similarities to any known protein. we generated specific antibodies against sars protein 3a by using a synthetic peptide (p2) corresponding to amino acids 261-274 of the putative protein. anti-p2 antibodies and the sera from sars patients could specifically detect the recombinant sars protein 3a expressed in esch ...	2004	15135062
severe acute respiratory syndrome: review and lessons of the 2003 outbreak.		2004	15155694
the spectrum of severe acute respiratory syndrome-associated coronavirus infection.	whether subclinical or atypical presentations of severe acute respiratory syndrome (sars) occur and whether clinical judgment is accurate in detecting sars are unknown. objectives: to describe the spectrum of sars coronavirus infection in a large outbreak and to compare diagnoses based on clinical judgment with the sars coronavirus test.	2004	15096332
summaries for patients. do some patients with sars have mild disease?		2004	15096361
severe acute respiratory syndrome (sars): a review.	severe acute respiratory syndrome (sars) is a newly emerged disease that rapidly spread around the world. the disease originated in southern china and a novel coronavirus (sars cov) has been implicated as the causative organism. the path this virus took to set up human infection remains a mystery, though preliminary data point to origins in an animal reservoir. nosocomial transmission of sars cov has been a striking feature in this epidemic. the clinical illness is similar to many acute respirat ...	2004	15139736
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 ...	2004	15140958
ph-dependent entry of severe acute respiratory syndrome coronavirus is mediated by the spike glycoprotein and enhanced by dendritic cell transfer through dc-sign.	the severe acute respiratory syndrome coronavirus (sars-cov) synthesizes several putative viral envelope proteins, including the spike (s), membrane (m), and small envelope (e) glycoproteins. although these proteins likely are essential for viral replication, their specific roles in sars-cov entry have not been defined. in this report, we show that the sars-cov s glycoprotein mediates viral entry through ph-dependent endocytosis. further, we define its cellular tropism and demonstrate that virus ...	2004	15140961
[sars, pandemic influenza, avian influenza: quest for missing link].		2004	15160886
bioinformatics analysis of sars coronavirus genome polymorphism.	we have compared 38 isolates of the sars-cov complete genome. the main goal was twofold: first, to analyze and compare nucleotide sequences and to identify positions of single nucleotide polymorphism (snp), insertions and deletions, and second, to group them according to sequence similarity, eventually pointing to phylogeny of sars-cov isolates. the comparison is based on genome polymorphism such as insertions or deletions and the number and positions of snps.	2004	15161495
organ distribution of severe acute respiratory syndrome (sars) associated coronavirus (sars-cov) in sars patients: implications for pathogenesis and virus transmission pathways.	we previously identified the major pathological changes in the respiratory and immune systems of patients who died of severe acute respiratory syndrome (sars) but gained little information on the organ distribution of sars-associated coronavirus (sars-cov). in the present study, we used a murine monoclonal antibody specific for sars-cov nucleoprotein, and probes specific for a sars-cov rna polymerase gene fragment, for immunohistochemistry and in situ hybridization, respectively, to detect sars- ...	2004	15141376
tissue distribution of ace2 protein, the functional receptor for sars coronavirus. a first step in understanding sars pathogenesis.	severe acute respiratory syndrome (sars) is an acute infectious disease that spreads mainly via the respiratory route. a distinct coronavirus (sars-cov) has been identified as the aetiological agent of sars. recently, a metallopeptidase named angiotensin-converting enzyme 2 (ace2) has been identified as the functional receptor for sars-cov. although ace2 mrna is known to be present in virtually all organs, its protein expression is largely unknown. since identifying the possible route of infecti ...	2004	15141377
re: to kf, tong jh, chan pk, et al. tissue and cellular tropism of the coronavirus associated with severe acute respiratory syndrome: an in-situ hybridization study of fatal cases. j pathol 2004; 202: 157-163.		2004	15141389
s protein of severe acute respiratory syndrome-associated coronavirus mediates entry into hepatoma cell lines and is targeted by neutralizing antibodies in infected patients.	the severe acute respiratory syndrome-associated coronavirus (sars-cov) causes severe pneumonia with a fatal outcome in approximately 10% of patients. sars-cov is not closely related to other coronaviruses but shares a similar genome organization. entry of coronaviruses into target cells is mediated by the viral s protein. we functionally analyzed sars-cov s using pseudotyped lentiviral particles (pseudotypes). the sars-cov s protein was found to be expressed at the cell surface upon transient t ...	2004	15163706
sars in the intensive care unit.	approximately 20% of patients with severe acute respiratory syndrome (sars) develop respiratory failure that requires admission to an intensive care unit (icu). old age, comorbidity, and elevated lactate dehydrogenase on hospital admission are associated with increased risk for icu admission. icu admission usually is late and occurs 8 to 10 days after symptom onset. acute respiratory distress syndrome occurs in almost all admitted patients and most require mechanical ventilation. icu admission i ...	2004	15142487
rapid identification of coronavirus replicase inhibitors using a selectable replicon rna.	a previously unknown coronavirus (cov) is the aetiological agent causing severe acute respiratory syndrome (sars), for which an effective antiviral treatment is urgently needed. to enable the rapid and biosafe identification of coronavirus replicase inhibitors, we have generated a non-cytopathic, selectable replicon rna (based on human cov 229e) that can be stably maintained in eukaryotic cells. most importantly, the replicon rna mediates reporter gene expression as a marker for coronavirus repl ...	2004	15166457
hiv protease inhibitor nelfinavir inhibits replication of sars-associated coronavirus.	a novel coronavirus has been identified as an etiological agent of severe acute respiratory syndrome (sars). to rapidly identify anti-sars drugs available for clinical use, we screened a set of compounds that included antiviral drugs already in wide use. here we report that the hiv-1 protease inhibitor, nelfinavir, strongly inhibited replication of the sars coronavirus (sars-cov). nelfinavir inhibited the cytopathic effect induced by sars-cov infection. expression of viral antigens was much lowe ...	2004	15144898