SARS病毒抗体间接免疫荧光检测方法的建立  

DEVELOPMENT OF IFA METHOD FOR DETECTING ANTIBODIES OF SARS CORONAVIRUS

作　　者：司炳银[1] 杨保安[1] 于曼[1] 刘洪[1] 吕富双[1] 韩伟国[1] 张雨[1] 石玉玲[2] 李林海[2] 秦鄂德[1] 祝庆余[1] 

机构地区：[1]军事医学科学院微生物流行病研究所 [2]广州军区广州总医院

出　　处：《解放军医学杂志》2003年第8期699-700,共2页Medical Journal of Chinese People's Liberation Army

摘　　要：目的　建立快速检测严重急性呼吸综合征 (SARS)患者血清中特异抗体的间接免疫荧光方法 ,为SARS的临床确诊提供参考依据。方法　用本实验室新分离的SARS冠状病毒BJ0 1和GZ0 1株感染Vero E6细胞 ,待 2 5 %细胞出现病变时 ,用胰蛋白酶消化细胞后以 2× 10 7/ml浓度 4 0 μl的细胞滴到 10孔镀膜的玻片上 ,CO2 温箱培养 4h后丙酮固定。利用制备的抗原片通过间接免疫荧光检测了从北京和广州地区采集的 15 4例临床诊断为SARS的患者和 14例非SARS呼吸系统疾病和 10 0份健康人血清。结果　成功地建立了检测SARS冠状病毒抗体的间接免疫荧光染色方法 ,并对 15 4例临床诊断为SARS的患者的血清标本进行检测 ;SARS冠状病毒抗体阳性 14 2例 ,阳性率为 92 3% ,而 14例非SARS呼吸系统疾病患者和 10 0名健康人血清检测均为阴性。结论　本方法具有良好的特异性和灵敏度 。Objective To develop a diagnostic test based on indirect immunofluorescence assay(IFA) to detect special antibodies in the serum of SARS patients, thus to provide a reference material for confirmation of the clinical diagnosis of SARS. Methods SARS coronavirus GZ01 and BJ01 strains isolated in our laboratory were used to infect Vero E6 cells. When CPE reached 25%, cells were trypsinized and transferred to 10 well slides in a quantity of 40μl with a cell density of 2×10 7 /ml. After 4 hour incubation at 37℃,the slides were fixed with acetone, and IFA was used to detect antibodies in serum samples, which were obtained from 154 SARS patients and 14 non SARS patients with respiratory disease, as well as 100 healthy volunteers. Results IFA method for detecting antibodies of SARS coronavirus was developed. Sera from one hundred and forty two out of 154 clinically diagnosed patients were IFA positive, with a positive rate of 92 3%. Sera from 14 non SARS patients with respiratory disease and 100 healthy persons were all IFA negative. Conclusion The IFA method we developed was sensitive and specific in detecting SARS antibodies in serum, and was a reliable test for laboratory diagnosis of SARS coronavirus.

关 键 词：严重急性呼吸综合征 冠状病毒 间接免疫荧光 

分 类 号：R563.802[医药卫生—呼吸系统]