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目的 研究SARS冠状病毒对不同组织的感染情况 ,了解SARS流行初期病毒的基因组序列。方法 用TrizolRNA抽提试剂提取因SARS病毒感染而死亡的 3例死者不同组织中的总RNA ,并逆转录为cDNA ,然后用针对SARS病毒不同基因区间的引物进行PCR扩增 ,并对其中 1例肺组织中病毒全序列进行了测定。结果 3例病例中有 2例仅在肺组织中扩增到病毒序列 ,另 1例除在肺组织中检测到病毒外 ,还在气管、肾、淋巴结及肝组织中检测到病毒的存在。病毒全序列分析结果显示 ,该序列全长为 2 976 0个碱基 ,具有典型的冠状病毒序列特征 ,除具有RNA聚合酶基因和S、E、M和N 4种结构蛋白基因外 ,还有另外 8个蛋白编码框。经与GenBank上登录的部分其他SARS冠状病毒全序列进行比对 ,发现该序列除存在少量的SNP外 ,还多出一段 2 9个核苷酸的序列 ,这段序列的存在完全改变了ORF1 0和ORF1 1两个蛋白编码框 ,使得在此终止的蛋白翻译能够继续进行下去 ,从而使ORF1 0和ORF1 1两个读框成为一个读框。结论 SARS病毒具有多组织嗜性 ;该序列有可能是病毒从动物传到人的原始序列
Objective To study the SARS-CoV infection in different tissues and to understand the genome sequence of the SARS-prevalent virus. Methods Trizol RNA extraction reagent was used to extract total RNA from different tissues of 3 deceased persons died of SARS virus infection and reverse transcribed into cDNA. Then, PCR was performed with primers targeting different SARS virus genes and 1 The complete sequence of the virus in lung tissue was determined. Results Of the 3 cases, 2 cases only amplified the virus sequence in the lung tissue. The other 1 case detected the virus in the trachea, kidney, lymph nodes and liver tissues in addition to the virus detected in the lung tissue. The results of full-length virus sequence analysis showed that the full-length sequence of the virus was 297600 bp with typical coronavirus sequences. In addition to the RNA polymerase gene and S, E, M and N four structural protein genes, Another 8 protein coding boxes. Comparing with other sequences of SARS coronavirus registered in GenBank, a sequence of 29 nucleotides was found in addition to a small number of SNPs. The presence of this sequence completely changed ORF10 And ORF1 1 two protein coding frames so that the protein translation terminating here can continue so that the two reading frames of ORF1 0 and ORF1 1 become a reading frame. Conclusion The SARS virus has multi-tissue tropism; this sequence may be the virus transmitted from animals to the human original sequence