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目的 分离和鉴定引起广东省非典型肺炎的病原体。方法 收集非典型肺炎患者的各种临床标本 ,用狗肾传代细胞 (MDCK)进行病原体分离 ,并运用血清学和分子生物学方法进行鉴定。结果 用MDCK细胞从非典型性肺炎患者标本中分离到病毒 ,用SARS病毒聚合酶基因的特异引物 ,通过巢式聚合酶链反应 (nestRT PCR) ,扩增出一个 2 79nt的片段 ,序列分析结果显示 ,该序列和目前已知的冠状病毒有 39%～ 6 5 %同源性 ,和来源于不同地区分离的SARS冠状病毒株 (中国北京、香港、台湾和德国、意大利等 )在同一个位置 (第 12个碱基 )仅有一个碱基不同 ,由a→t。间接免疫荧光抗体检测显示 ,该病毒能够和非典型肺炎患者的恢复期血清呈特异的抗体反应。结论 这种新的冠状病毒是引起广东省非典型肺炎的病原 ,且能够用MDCK细胞进行分离
Objective To isolate and identify the pathogens causing atypical pneumonia in Guangdong Province. Methods Various clinical specimens of patients with atypical pneumonia were collected. Pathogen isolation was carried out using MDCK in dogs and identified by serological and molecular biological methods. Results The virus was isolated from the specimens of patients with atypical pneumonia using MDCK cells. A 2 79 nt fragment was amplified by nestRT PCR using specific primers of the SARS virus polymerase gene. The results of sequence analysis The sequence showed 39% ~ 65% homology with the known coronavirus at the same location as the SARS coronavirus strains (Beijing, Hong Kong, Taiwan and Germany, Italy, etc.) isolated from different regions (12th base) differs only by one base from a → t. Indirect immunofluorescence antibody test showed that the virus was able to show specific antibody response to convalescent sera of patients with atypical pneumonia. Conclusion This new coronavirus is the causative agent of atypical pneumonia in Guangdong province and can be isolated by MDCK cells