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目的了解海岛地区病毒性腹泻病原中诺如病毒的流行状况,对优势流行株进行基因序列分析。方法对舟山市监测医院疑似病毒性腹泻粪便样本经酶联免疫吸附法(ELISA)筛选轮状病毒,对阴性样本采用逆转录聚合酶链反应(RT-PCR)扩增诺如病毒RNA聚合酶与主要衣壳蛋白基因,PCR扩增产物进行测序反应,序列结果与GenBank已公布的序列进行比对,测其同源性;并绘制成系统发生树进行进化分析。结果在35份诺如病毒扩增阳性样本中挑选病毒载量最高的Hu/Zhejiang/9/2006/CHN株进行测序,并与Hunter株、Farming Hill株等进行比对并绘制系统发生树。Hu/Zhejiang/9/2006/CHN株属于诺如病毒GⅡ-4群,与Hunter株和Lanzhou株形成的分支较接近,RNA聚合酶有部分序列与这2株完全相同。结论舟山市(海岛地区)病毒性腹泻病原中存在诺如GⅡ-4群,采用GDD基序分析有利于更好地发现新的变异株的存在,为病毒性腹泻的病原学和基因学研究提供依据。
Objective To understand the prevalence of norovirus in the viral diarrheal pathogen in the island area and to analyze the gene sequence of dominant epidemic strains. Methods Rotavirus was screened by enzyme-linked immunosorbent assay (ELISA) in the suspicious viral diarrhea stool samples of Zhoushan City Surveillance Hospital. Norovirus RT-PCR was used to amplify norovirus RNA polymerase against negative samples The major capsid protein gene and PCR amplification products were sequenced. The sequence was compared with the published sequences in GenBank to test the homology. The phylogenetic tree was also constructed for phylogenetic analysis. Results The 35 strains of Huo Zhejiang / 9/2006 / CHN strains with the highest viral load were selected for sequencing and compared with those of Hunter strain and Farming Hill strain. The phylogenetic tree was constructed. The strain Hu / Zhejiang / 9/2006 / CHN belonged to the Norovirus GⅡ-4 group, which was closer to the branch formed by the Hunter strain and Lanzhou strain. The partial sequence of RNA polymerase was exactly the same as that of the two strains. Conclusion Norovirus GⅡ-4 population exists in the pathogen of viral diarrhea in Zhoushan (island area). GDD motif analysis is helpful to discover new mutant strains better and provide etiological and genetic studies on viral diarrhea in accordance with.