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Virologica Sinica, 18 (6) : 544-547, 2003
Research Article
Isolation and Identification of SARS.CoronaviFUS
ZhongshahMedical College,Sun Yat·sen University,Guangzhou 510080,China
2.FirstMilitaryMedical University,Guangzhou 5105l5.China
Abstract:To isolate and identify the etiologic agent of severe acute respiratory syndrome(SARS),
the throat swabs an d gargle of patients with SARS were collected.We inoculated the specimens onto a
number of cell lines including Vero,Vero E6,M DCK,Hela,Hep一2,HP an d HEL cells.The isolates
were identified by thin—section electron.microscopy,indirect.immunofluorescence,neutralization test,
RT—PCR an d nucleotide sequence an alysis.Using cell culture techniques.a coronavirus was isolated
from patient with SARS.Four celllines,Vero,Vero E6,HP an d MDCK cells,showed cytopathic efec t.
1]he infected cells showed strong cytoplasmi c an d membran ous staining with a convalescent—phase
seruln from the patients with SARS in an indirect—immunofluorescence staining.The cytopathic efect
of the viruses clould been neutralized by convalescent serum.Under elec tron—microscopy,a large
num be r of coronavirus—like particles were obsewed in the infected cells.W ith specific primers.the
specific cDNA fragments were amplified by reverse transcription—polymerase chain reaction(RT—PCR1
from infected cels an d the nucleotide sequence analysis showed that homology of the amplified
fragm ents to the SARS—coronavirus(SARS·CoV)previously reported was 100%.A SARS—CoV Was
isolated from patients with SARS.T}liS virus was closely associated with SARS.
  Published online: 25 Dec 2003
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