雷玥, 庄志超, 田宏, 董兴华, 李晓燕. 2020 年天津市宁河区急性胃肠炎聚集疫情病原体轮状病毒全基因组序列分析[J]. 疾病监测, 2021, 36(5): 417-422. DOI: 10.3784/jbjc.202012020407
引用本文: 雷玥, 庄志超, 田宏, 董兴华, 李晓燕. 2020 年天津市宁河区急性胃肠炎聚集疫情病原体轮状病毒全基因组序列分析[J]. 疾病监测, 2021, 36(5): 417-422. DOI: 10.3784/jbjc.202012020407
Lei Yue, Zhuang Zhichao, Tian Hong, Dong Xinghua, Li Xiaoyan. Whole genome analysis on rotavirus isolated in outbreak in Ninghe district of Tianjin, 2020[J]. Disease Surveillance, 2021, 36(5): 417-422. DOI: 10.3784/jbjc.202012020407
Citation: Lei Yue, Zhuang Zhichao, Tian Hong, Dong Xinghua, Li Xiaoyan. Whole genome analysis on rotavirus isolated in outbreak in Ninghe district of Tianjin, 2020[J]. Disease Surveillance, 2021, 36(5): 417-422. DOI: 10.3784/jbjc.202012020407

2020 年天津市宁河区急性胃肠炎聚集疫情病原体轮状病毒全基因组序列分析

Whole genome analysis on rotavirus isolated in outbreak in Ninghe district of Tianjin, 2020

  • 摘要:
      目的  分析2020年天津市宁河区一起急性胃肠炎聚集疫情的病原A组轮状病毒(RVA)的全基因组序列,调查原因及防控措施。
      方法  对送检轮状病毒阳性粪便标本核酸进行提取,经real-time PCR进一步确证;构建DNA测序文库,采用二代测序法进行测序,合格序列拼接为11条基因片段。 对轮状病毒重要VP7、VP4和NSP4等序列进行系统进化和重要抗原表位变异分析。
      结果  经确认4份样本为RVA阳性,二代测序成功获得4株轮状病毒全基因序列。 4株RVA相似性为99%以上,所获毒株均为Wa-like基因组群,G9-P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1。 亲缘关系树分析所有毒株的VP7基因属于G9-Ⅵ,VP4基因属于P[8]-3亚型。 基因序列进一步揭示了VP7和VP4抗原表位存在氨基酸位点替代现象。 并且在非结构蛋白NSP4基因的Ⅰ型和Ⅱ型抗原表位上存在氨基酸位点的改变。 其余结构蛋白和非结构蛋白均相对保守。
      结论  G9-Ⅵ亚型、P[8]-3亚型RVA是本次疫情的病原体,本次所获得的4株轮状病毒与现流行毒株变异较小,但应加强轮状病毒全基因组的进化监测,为掌握其流行趋势的变化和开展有效的疫情防控提供科学数据。

     

    Abstract:
      Objective  To understand the whole genome characteristics of rotavirus A strains isolated from acute gastroenteritis cases in an outbreak in Ninghe district of Tianjin in 2020.
      Methods  The viral nucleic acids of rotavirus isolated from positive stool samples collected in Ninghai were extracted for real-time PCR confirmation, the sequencing library were constructed following the instructions, the gene sequencing was conducted by the second generation sequencing method, and the sequencing data were analyzed by phylogenetic software, and 11 gene fragments full-genome sequences were analyzed by homology and phylogenetic analyses.
      Results  Complete gene sequences of 4 strains of rotavirus were obtained successfully by the second generation sequencing method. All the strains shared >99% homology and showed the Wa-like constellation G9-P[8]-I1-R1-C1-M1-A1-N1-T1-E1-H1. Furthermore, phylogenetic analysis revealed that the VP7 genes belonged to lineage G9-VI and the VP4 sequences belonged to lineage P[8]-3. Amino acid substitutions were observed in the antigenic regions of the VP7 and VP4 gene. The nonstructural protein 4 (NSP4) genes contained multiple substitutions in antigenic sites Ⅰ and Ⅱ. The other NSP genes were conserved.
      Conclusion  The study revealed that rotavirus G9-VI and P[8]-3 were the pathogens of this outbreak. The variations of the 4 strains were minor compared with the circulating strain. The continuous surveillance and analysis of the whole genomic constellation of circulating rotavirus strains needs to be strengthened to provide evidence for the prevention and control of rotavirus infection.

     

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