摘要
目的 研究SARS患者中多病原感染状况，测定并分析新分离呼肠病毒基因组序列，从病毒的分子生物学角度研究其分类学位置。方法 SARS患者咽拭子标本经处理后接种Hep-2细胞，提取分离到的病毒RNA，以随机引物逆转录PCR扩增，PCR产物经克隆、测序，将其核苷酸序列及推断的氨基酸序列与GenBank（数据库中基因进行比较分析并建立系统进化树。结果 从4份SARS患者咽拭子标本中分离到3株呼肠病毒，部分基因片段测序结果显示为同一病毒，其病毒基因与呼肠病毒1、2、3型有较高的同源性，尤其是与呼肠病毒1、3型，相应片段的核苷酸序列同源性达82％～92％，推断相应氨基酸序列同源性高达94％～99％，而与其他病毒不存在有意义的同源性；系统进化树分析显示新分离呼肠病毒属于呼肠病毒科呼肠病毒属，且可能为哺乳动物呼肠病毒中的新血清型。结论 SARS患者咽拭子标本中存在呼肠病毒，新分离呼肠病毒核苷酸和氨基酸序列与呼肠病毒1、3型同源性最高，由于尚未得到决定病毒血清型的S1片段基因序列，故该病毒的确切分型有待进一步确定，与SARS患者病情的关系有待进一步研究。
Objective To clarify whether patients with SARS may harbour a reovirus and identify a new isolated reovirus using molecular assays. Methods Four throast-swab specimens were received from 4 patients with SARS and tested by using virus-isolation techniques and reverse transcription-polymerase chain reaction （RT- PCR） ,in an attempt to obtain the sequence of the new isolated reovirus. Results A novel reovirus was isolated from 4 patients who met the case definition of SARS. Cytopathological features were noted in Hep-2 cells inoculated with throat-swab specimen. Sequence analysis showed that the newly isolated reovirus shared 82 %-92 % nucleotide identity with reovirus serotype 1,2 and 3.Amino acid identity was 94%-99%. Phylogenetic analysis of the new isolated reovirus showed that it was probably a new serotype of mammalian orthoreovirus. Conclusion There is reovirus in the throat-swab specimens from patients with SARS. The newly isolated reovirus shared high identity with reovirus serotype 1,2 and 3. But the relationship between new isolated reovirus and SARS is unclear.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
北大核心
2006年第4期373-375,共3页
Chinese Journal of Experimental and Clinical Virology
基金
国家自然科学基金项目(30471555)
