广州地区传染性非典型肺炎病原体分离结果和特性分析  

Characteristics of the pathogenic isolates from the SARS patients in Guangzhou

作　　者：何丽娟[1] 狄飚[1] 周端华[1] 杨卫路[1] 邱季春[1] 陈芳[1] 周秀珍[1] 吴新伟[1] 高阳[1] 鲁恩洁[1] 张玮[1] 

机构地区：[1]广州市疾病预防控制中心,510800

出　　处：《华南预防医学》2003年第3期32-33,T001,共3页South China Journal of Preventive Medicine

摘　　要：目的　分析从传染性非典型肺炎 (SARS)患者标本中分离到的病原体及其特性。方法　将采集的SARS患者的咽漱液、咽拭子、尸解组织或尸解组织液分别接种到人胚肺细胞 (HL)、狗肾细胞 (MDCK)和人咽喉癌上皮细胞 (Hep 2 ) ,37℃、34℃二氧化碳培养箱培养。样本原液、细胞培养阳性的上清液分别接种 9日龄鸡胚羊膜腔、尿囊腔各 0 2ml,3d后分别收集尿液、羊膜液。用 1 %鸡红细胞、0 75 %豚鼠红细胞、人“O”型红细胞和鹌鹑红细胞做血细胞凝集试验。病变细胞用 0 2 %豚鼠红细胞做红细胞吸附试验。制备的病毒抗原片用恢复期患者血清做免疫荧光抗体检测 ,尸解肺组织作电镜检测 ,分离的病原体用RT PCR方法检测 ,用ELISA方法检测患者急性期、恢复期双份血清IgG抗体。结果　 75份样本细胞分离到 1 3株病原体 ,分离阳性率为 1 7 3%。病原体在 34℃环境中生长繁殖较 37℃好 ,而且HL较Hep 2敏感 ,MDCK不敏感。病原体可使人“O”型红细胞和鹌鹑红细胞凝集 ,而不能使鸡红细胞和豚鼠红细胞凝集。病变细胞用免疫荧光试验检测病人的血清 ,结果呈阳性。尸解肺组织电镜检查找到一种冠状病毒样颗粒。分离的病原体用RT PCR方法检测可分别扩增出冠状病毒的特异基因片段。用ELISA方法检测患者恢复期血清IgG抗体阳性率为 85 7%Objective To investigate the pathogenic isolates from clinical specimens of the SARS patient. Method Throat swabs, throat washings and autopsy tissues collected from the SARS patients were inoculated into cell lines of HL、MDCK and Hep 2 for culture under 37℃ and 34℃. The specimens and the culture medium of the cell with cytopathic effect also inoculated into the amniotic and allantoid cavities of 9 day old embryonated eggs. The urines and amnia fluids collected 3 days later to undergo hemagglutination test(HA) with 1% chicken, 0.75% guinea pig red blood cells, human “O” type red blood cell and the partridge red blood cells. The cells with cytopathic effect were tested through absorbed test with 0.2% guinea pig red blood cells. Immunofluorenscent stain was performed to test the antibody of the patient's convalescent phase serum. The lung tissues from the SARS bodies were examined by the electronic microscope and the isolated pathogen were identified through RT PCR. Results A novel corona virus isolated from 13 of 75 (17.3%) clinical specimens and autopsy tissues were confirmed by the electronic microscope examination and RT PCR. The virus could not grow in the MDCK cell line but could grow in the Hep 2. It grew in the condition of 34℃ better than in 37 ℃. The surface of the virus is of agglutinin which is capable of introducing agglutination of human “O ' type red blood cell and the partridge red blood cells. But could not show the same effect in that of the chicken and guinea pig. The new isolate demonstrate a virus specific serologic response through immunofluorescent antibody test. About 85.7% (48/56) of dual serum IgG were positively detected by ELISA. Conclusion A novel corona virus is associated with this outbreak of SARS in Guangzhou recently.

关 键 词：广州地区 传染性非典型肺炎 严重急性呼吸综合征 冠状病毒 病原体分离 

分 类 号：R563.1[医药卫生—呼吸系统]