Isolation and Identification of SARS．CoronaviFUS

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．