【摘要】 为查找引起广州地区流行的严重急性呼吸综合征(SARS)的病原体,采集患者漱口液及尸解标本,用组织培养法接种人胚肺细胞、MDCK细胞、Hep-2细胞和鸡胚分离病毒,用间接免疫荧光法检测患者恢复期血清IgG抗体,确定分离的病原是SARS的主要病因,再用套式RT-PCR、免疫电镜法鉴定病原。结果用人胚肺、Hep-2细胞在75份漱口液和3例尸解组织中分离出13株病原体,经套式RT-PCR扩增出110bp的特异产物,经测序证实为冠状病毒。制备冠状病毒的抗原,检测30份SARS病人恢复期血,其中26份血清IgG抗体阳性。同时检测30份普通发热病人血清作对照,IgG抗体全部阴性。由此证明,经组织培养分离到的病原体是引起SARS的致病因子,用分子生物学方法测序后证实为冠状病毒。更多还原

【Abstract】 To investigate the pathog en that caused the epidemic of severe acute respirat ory syndrome (SARS )in Guangzhou region,the clinical specimens including oropharyng eal washings and autopsy specimens were collected and inoculated onto cell line monolayer, including human embryonic lung, MDCK, Hep-2 and chic ken embro for virus isolation. Pati ents’serum IgG antibody was als o detected by indirect immunofluor escence assay(IFA)during the conva lescent phase.The isolated pathoge n was further detected by nested r everse transcription-polymerase ch ain reaction(RT-PCR)and immunoelec tron microscopy.Thirteen pathogens were successfully isolated in huma n embryonic lung and Hep-2 cell fr om 75 oropharyngeal washings and 3 autopsy specimens,and a specific s equence of 110bp product was obtai ned by the nested RT-PCR amplifica tion procedure.The product was con firmed as coronavirus by DNA seque ncing.The prepared coronavirus ant igen was used to test thirty SARS patients’blood specimens during co nvalescent phase.Twenty six among thirty blood specimens showed IgG antibody positive,by the same time ,all thirty control sera(from common feverish patients)were all nega tive for SARS IgG antibody.By immuno electron microscopy,coronavirus an d chlamydial particles both were f ound in autopsy lung tissue combin ed with SARS patient’s convalescen t serum.Therefore,pathogen isolate d from tissue culture is believed to be responsible for SARS.The seq uencing data of the isolated patho gen confirms that the virus is cor onavirus.更多还原