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病原学研究一种与SARS有关的新型冠状病毒
N
Engl J Med 2003 Apr 16; [epub ahead of print ]
A Novel Coronavirus Associated with Severe Acute Respiratory
Syndrome.
Ksiazek TG, Erdman
D, Goldsmith CS, Zaki SR, Peret T, Emery S, Tong S, Urbani
C, Comer JA, Lim W, Rollin PE, Dowell SF, Ling AE, Humphrey
CD, Shieh WJ, Guarner J, Paddock CD, Rota P, Fields B, DeRisi
J, Yang JY, Cox N, Hughes JM, LeDuc JW, Bellini WJ, Anderson
LJ.
Background A worldwide
outbreak of severe acute respiratory syndrome (SARS) has been
associated with exposures originating from a single ill health
care worker from Guangdong Province, China. We conducted studies
to identify the etiologic agent of this outbreak. Methods
We received clinical specimens from patients in six countries
and tested them, using virus isolation techniques, electron-microscopical
and histologic studies, and molecular and serologic assays,
in an attempt to identify a wide range of potential pathogens.
Results No classic respiratory or bacterial respiratory pathogen
was consistently identified. However, a novel coronavirus
was isolated from patients who met the case definition of
SARS. Cytopathological features were noted microscopically
in Vero E6 cells inoculated with a throat-swab specimen. Electron-microscopical
examination of cultures revealed ultrastructural features
characteristic of coronaviruses. Immunohistochemical and immunofluorescence
staining revealed reactivity with group I coronavirus polyclonal
antibodies. Consensus coronavirus primers designed to amplify
a fragment of the polymerase gene by reverse transcription-polymerase
chain reaction (RT-PCR) were used to obtain a sequence that
clearly identified the isolate as a unique coronavirus only
distantly related to previously sequenced coronaviruses. With
specific diagnostic RT-PCR primers we identified several identical
nucleotide sequences in 12 patients from several locations,
a finding consistent with a point source outbreak. Indirect
fluorescent antibody tests and enzyme-linked immunosorbent
assays made with the new coronavirus isolate have been used
to demonstrate a virus-specific serologic response. Preliminary
studies suggest that this virus may never before have infected
the U.S. population. Conclusions A novel coronavirus is associated
with this outbreak, and the evidence indicates that this virus
has an etiologic role in SARS. The name Urbani SARS-associated
coronavirus is proposed for the virus.
Notice: Because
of possible public health implications, this article was published
at www.nejm.org on April 10, 2003. It will appear in the May
15 issue of the Journal. Click on "PDF of this article"
for the full text. Copyright 2003 Massachusetts Medical Society
从SARS患者中鉴定新型冠状病毒
N
Engl J Med 2003 Apr 10; [epub ahead of print]
Identification
of a Novel Coronavirus in Patients with Severe Acute Respiratory
Syndrome.
Drosten C, Gunther
S, Preiser W, Van Der Werf S, Brodt HR, Becker S, Rabenau
H, Panning M, Kolesnikova L, Fouchier RA, Berger A, Burguiere
AM, Cinatl J, Eickmann M, Escriou N, Grywna K, Kramme S, Manuguerra
JC, Muller S, Rickerts V, Sturmer M, Vieth S, Klenk HD, Osterhaus
AD, Schmitz H, Doerr HW.
Background The
severe acute respiratory syndrome (SARS) has recently been
identified as a new clinical entity. SARS is thought to be
caused by an unknown infectious agent. Methods Clinical specimens
from patients with SARS were searched for unknown viruses
with the use of cell cultures and molecular techniques. Results
A novel coronavirus was identified in patients with SARS.
The virus was isolated in cell culture, and a sequence 300
nucleotides in length was obtained by a polymerase-chain-reaction
(PCR)-based random-amplification procedure. Genetic characterization
indicated that the virus is only distantly related to known
coronaviruses (identical in 50 to 60 percent of the nucleotide
sequence). On the basis of the obtained sequence, conventional
and real-time PCR assays for specific and sensitive detection
of the novel virus were established. Virus was detected in
a variety of clinical specimens from patients with SARS but
not in controls. High concentrations of viral RNA of up to
100 million molecules per milliliter were found in sputum.
Viral RNA was also detected at extremely low concentrations
in plasma during the acute phase and in feces during the late
convalescent phase. Infected patients showed seroconversion
on the Vero cells in which the virus was isolated. Conclusions
The novel coronavirus might have a role in causing SARS.
Notice: Because
of possible public health implications, this article was published
at www.nejm.org on April 10, 2003. It will appear in the May
15 issue of the Journal. Click on "PDF of this article"
for the full text. Copyright 2003 Massachusetts Medical Society
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