临床诊断的SARS患者鼻咽拭子采集、SARS冠状病毒分离及鉴定

Isolation and Identification of SARS -Coronavirus in Nasal and Throat Swabs Collected from Clinically Diagnosed SARS Patients

目的从临床诊断的严重急性呼吸综合征(severeacuterespiratorysyndrome,SARS)患者鼻咽拭子标本分离并鉴定SARS冠状病毒。方法SARS患者鼻咽拭子标本接种于含Vero及MRC-5单层细胞的24孔培养板中。分离物使用血清学方法、电镜及基因测序进行鉴定。结果2003年4月～5月北京协和医院共采集79例临床诊断SARS患者的158份鼻咽拭子标本。3例接种Vero细胞孔的鼻拭子标本发现细胞病变效应(cytopathiceffect,CPE)。用分离物感染细胞做为抗原,免疫荧光血清学发现SARS患者急性期和恢复期血清标本存在SARS冠状病毒抗体(IgG和IgM)阳转和/或4倍及以上升高。分离物上清液负染电镜观察到冠状病毒。基因测序证实3株分离物是SARS冠状病毒。结论3株自79例SARS患者鼻咽拭子标本的病毒分离物是SARS冠状病毒。

To isolate and identify SARS-coronavirus in nasal and throat swabs collected from clinically diagnosed severe acute respiratory syndrome (SARS)patients.Methods Nasal and throat swab specimens were inoculated onto well of24-well plate containing confluent monolayers of Vero and MRC-5cells.Isolates were identified with serology,electron microscopy and genome sequence.Results One hundred and fifty-eight nasal and throat swabs specimens from79SARS patients in Peking Union Medical College Hospital between April and May,2003were cultured for SARS-coronavirus.Cytopathic effect (CPE)was found in three nasal swab specimens inoculated in Vero cells.Acute and convalescent phase serum specimens collected from SARS patients were found with seroconversions and/or a fourfold or greater rises in indirect fluorescence antibodies(IgG and IgM)titers when the3isolates(infected Vero cells)were used as antigen.Coronavirus was observed in the culture supernatant by negative-stain electron microscopy.Genome sequence confirmed the isolates were SARS-coronavirus.Conclusions The3isolates from nasal and throat swabs samples collected from79clinically diagnosed SARS patients were SARS coronavirus.