摘要
目的 应用人咽喉上皮癌细胞 (Hep 2 )对严重急性呼吸综合征 (SARS)病例标本进行病原体检测与研究 ,为该病的预防与控制提供依据。方法 采用Hep 2细胞培养法 ,对 2 0 0 3年 1～ 2月广东省收集的SARS病例的咽拭子等标本分离致病病原体 ,并应用电镜形态学、逆转录 多聚酶链反应 (RT PCR)扩增部分基因进行序列分析 ,并对分离的病原体进行研究。结果 132例SARS病例的 31份咽拭子、10 0份痰液和 1份尸检肺组织标本中有 73份标本致单层细胞出现“蚀斑病变” ,细胞病变阳性率为 5 5 .3% ,其中 4 8h阳性率为 36 .4 %。采用巢式PCR(Nested PCR)对其中 2 5份出现“蚀斑病变”细胞培养物进行检测 ,能扩增出冠状病毒的特异片段 ;对中 2 18、中 2 19等 6份Nested PCR产物进行基因序列分析 ,结果显示其核苷酸序列与国内外公布的结果一致 ,氨基酸序列与已知的人或动物冠状病毒的同源性在 5 8%～ 76 %之间。通过电镜在SARS病例标本 (中 2 10 )病变细胞及 1份尸检肺组织标本中观察到衣原体样颗粒和病毒样颗粒。结论 从接种SARS病例标本的Hep 2细胞病变培养物中证实有冠状病毒。
Objective Epithelial Hep-2 (Human larynx carcinoma) cell line was applied to isolate and to identify the pathogen of SARS, in order to provide evidence for SARS prevention and control. Methods The samples collected from SARS patients from January to February in 2003 were inoculated into Hep-2 cell culture. The isolates were studied through electronic microscopic, RT-PCR and partial gene sequence analysis. Results Seventy-three samples out of 132 samples from 132 SARS cases (31pharynx swabs, 100 sputums and 1 lung tissues) showed positive cytopathic effect in monolayer cells. The positive rate was 55.3% and the rate within 48 hours was 36.4%. Specific gene fragments of corona virus were obtained by nested-PCR from 25 positive cytopathic cell cultures and their sequence analyses correspond with the SARS-CoV reported before. The homogeneity of amino acid to the previously known human or animal corona virus approached to 58%-76%. Both Chlamydia-like and virus-like particles were found in a cytopathic cell and an autopsy lung sample by electron microscopy. Conclusion The corona virus RNA was detected in Hep-2 cytopathic cell culture inoculated with bio-samples from SARS case.
出处
《华南预防医学》
2003年第5期9-13,共5页
South China Journal of Preventive Medicine
