摘要
目的 利用冠状病毒全基因组芯片检测SARS患者血、便和痰标本中的冠状病毒核酸 ,为SARS病毒核酸的诊断提供线索。方法 提取SARS患者血、便和痰样本中的冠状病毒RNA ,经全基因组随机反转录和PCR扩增标记成荧光探针 ,与SARS病毒全基因组芯片进行杂交 ,同时用体外培养的病毒RNA和从健康人分离的白细胞的RNA分别做阳性和阴性对照 ,杂交后的芯片用Axon4 10 0A扫描仪扫描和分析。结果 SARS患者的 3种标本中都能检测到冠状病毒核酸的存在 ,与整个病毒基因组比较大都为一些不连续的片断。其中痰和便标本中的基因谱较为相似 ,并且发现编码S1蛋白的核酸在多数痰和便标本中是连续的。结论 冠状病毒全基因组芯片能够检测出SARS患者血、便和痰标本中的冠状病毒核酸 ,同时能够监测不同样本中该病毒全基因组变化的特点 。
Objective The nucleic acid of coronavirus in blood, feces and sputum specimens obtained from SARS patients was surveyed using whole genomic gene chip of coronavirus. The purpose of the study was to look for some molecular information for the diagnosis of coronavirus infection. Methods Total RNAs of coronavirus in the blood, feces and sputum of SARS patients were extracted and reversely transcripted using 6 random primers which were respectively labeled by fluorescent dye Cy5 through PCR reaction respectively. Then the three groups of labeled cDNA were hybridized with the whole genomic gene chip of coronaviruses. Simultaneously, probes from cultured coronaviruses and blood of healthy persons were used as positive and negative controls. The Axon 4100A machine was used to scan and analyze the signals on the hybridized chips. Results the nucleic acid of coronaviruses was detected in all the three specimens, and they were less discontiguous compared with the whole genomic sequence of coronavirus. The gene profile of coronaviruses in feces and sputum specimens was similar. In addition, the sequence coding S1 protein was more continuous in the majority of specimens of feces (48 of 51) and sputum (8 of 13) than others. It was also found that when the SARS patients recovered from the disease, the continuity of S1 sequence changed weakly in the fecal specimens. Conclusion The nucleic acid of coronavirus in blood, feces and sputum specimens of SARS patients can be detected using whole genomic gene chip of coronavirus. The whole genomic changes in coronaviruses in different specimens can also be observed with this chip
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2003年第11期998-1000,共3页
Medical Journal of Chinese People's Liberation Army
