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目的 探索 2 0 0 2年末暴发于广东等地的非典型肺炎的病原。方法 对来自广东、山西、四川省和北京市的 7例非典型肺炎死亡病人的尸体解剖标本进行超薄切片电镜观察 ,将病人肺组织提取物接种于 2 93细胞 (人胚肾转化细胞系 ) ,分离病原 ,对病人组织中和分离的病原进行免疫学鉴定。结果 超薄切片电镜观察 ,在死亡患者的肺 ( 7例 )、脾 ( 2例 )、肝 ( 2例 )、肾 ( 3例 )和淋巴结 ( 1例 )中均观察到衣原体样包涵体和网质小体、中间体、原生小体颗粒的存在 ,并在 3例病人肺组织中同时见到了冠状病毒样颗粒和未知的基质样结构。用RT PCR方法从 2例病人肺组织中扩增出冠状病毒RNA聚合酶基因片段。将病人肺组织提取物接种于 2 93细胞进行培养 ,取第 2代细胞作电镜观察 ,也发现衣原体样颗粒和包涵体的存在。在病人组织和培养细胞中见到的衣原体样颗粒均不能与衣原体属特异性抗体和抗肺炎衣原体、鹦鹉热衣原体抗体反应 ,提示可能为新的衣原体样因子。结论在死亡病人组织和培养细胞中发现的衣原体样和冠状病毒样颗粒 ,可能是广东等地暴发的非典型肺炎的主要病原。
Objective To explore the etiology of atypical pneumonia outbreak in Guangdong and other places in late 2002. Methods The autopsy specimens from 7 dead SARS patients from Guangdong, Shanxi, Sichuan and Beijing were observed by ultra-thin section electron microscopy. The lung tissue extracts were inoculated into 293 cells (human embryonic kidney cell line) , Isolating the pathogen, immunologically identifying the pathogen in the patient’s tissue and isolating it. Results Ultrathin section electron microscopy showed that chlamydial inclusion bodies and nets were observed in the lungs (7 cases), spleen (2 cases), liver (2 cases), kidney (3 cases) and lymph nodes (1 case) Plastids, intermediates, and primary body particles, and coronavirus-like particles and unknown matrix-like structures were seen in the lung tissue of 3 patients. The coronavirus RNA polymerase gene fragment was amplified from the lung tissue of 2 patients by RT PCR. The patient’s lung tissue extracts were inoculated on 293 cells for culture, and the second generation cells were subjected to electron microscopy to observe the presence of chlamydia-like particles and inclusion bodies. Chlamydia-like particles seen in both patient tissue and cultured cells failed to react with Chlamydia-specific antibodies and anti-Chlamydia pneumoniae and Chlamydia psittaci antibodies, suggesting a possible new chlamydial-like factor. Conclusions Chlamydia-like and coronavirus-like particles found in the tissue of the deceased patient and in the cultured cells may be the major causative agent of the outbreak of SARS in Guangdong and other places.