血、便、痰和尿标本中SARS冠状病毒核酸的检测和病毒基因谱的初步建立

The detection of coronavirus nucleic acid and construction of its gene profile in blood、feces、sputum and urine specimens from SARS patient

  • 摘要: 目的:利用冠状病毒全基因组芯片检测SARS患者血、便、痰和尿标本中的冠状病毒核酸并试图建立其基因谱。方法:提取SARS患者血、便、痰和尿样本中的冠状病毒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和从健康人血液中提取的RNA做对照。结果:SARS患者的4种标本中都能检测到冠状病毒核酸的存在,与整个病毒基因组相比,大都为一些不连续的片断。其中尿、痰和便标本中的基因谱较为相似,另外,发现编码S1蛋白的核酸在一些标本中是连续的,具体为23份血标本中有2份是连续的,13份痰标本中有8份是连续的,51份便标本中有48份是连续的,2份尿标本中都是连续的。结论: 冠状病毒全基因组芯片能够检测出SARS患者血、便、痰和尿标本中的冠状病毒核酸,并能建立各来源标本中病毒核酸的基因谱。

     

    Abstract: Objective:To detect the coronavirus‘s nucleic acid in blood、feces、sputum and urine specimens from SARS patient and construt the corresponding gene profile of coronavirus in those specimen using whole genomic gene chip of coronavirus. Methods:Total RNAs of coronavirus,which come from blood,feces,sputum and urine of SARS patients,were extracted,reverse transcripted using random primers and labeled to be probes by fluorescent dye through PCR reaction respectively.Then the four groups probes were hybridized with the whole genomic gene chip of coronavirus.Simltaneitily,probes from cultured coronavirus and healthy blood were used to be positive and negative control.Results:the nucleic acid of coronavirus was detected in all the four specimens,but the nucleic acid was less integrated compare with the whole genomic sequence of the coronavirus ,and the gene profile of the coronavirus which come from feces,urine and sputum specimens was similar.In addation,the sequence that code the S1 protein was more continuous in some specimens compare with other sequences,those specimens include 2of 23blood samples,8 of 13sputum samples,48 of 51 samples and 2 of 2 urine samples.Conclusion:The coronavirus‘s nucleic acid in blood、feces、sputum and urine specimens of SARS patient can be detected using whole genomic gene chip of coronavirus. and the corresponding gene profile of coronavirus in those specimen can be constructed also.

     

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