在中国分离的 G9P[8]-E2 型轮状病毒的重配和基因组分析。
Reassortment and genomic analysis of a G9P[8]-E2 rotavirus isolated in China.
机构信息
Department of Biosciences, Faculty of Sciences, Universiti Teknologi Malaysia, Johor Bahru, 81310, Malaysia.
NHC Key Laboratory of Medical Viruses and Viral Diseases, Institute of Viral Disease Prevention and Control, National Health Commission, Chinese Centre for Disease Control and Prevention, Beijing, 102206, China.
出版信息
Virol J. 2023 Jun 22;20(1):135. doi: 10.1186/s12985-023-02064-5.
OBJECTIVE
To isolate a prevalent G9P[8] group A rotavirus (RVA) (N4006) in China and investigate its genomic and evolutionary characteristics, with the goal of facilitating the development of a new rotavirus vaccine.
METHODS
The RVA G9P[8] genotype from a diarrhea sample was passaged in MA104 cells. The virus was evaluated by TEM, polyacrylamide gel electrophoresis, and indirect immunofluorescence assay. The complete genome of virus was obtained by RT-PCR and sequencing. The genomic and evolutionary characteristics of the virus were evaluated by nucleic acid sequence analysis with MEGA ver. 5.0.5 and DNASTAR software. The neutralizing epitopes of VP7 and VP4 (VP5* and VP8*) were analyzed using BioEdit ver. 7.0.9.0 and PyMOL ver. 2.5.2.
RESULTS
The RVA N4006 (G9P[8] genotype) was adapted in MA104 cells with a high titer (10 PFU/mL). Whole-genome sequence analysis showed N4006 to be a reassortant rotavirus of Wa-like G9P[8] RVA and the NSP4 gene of DS-1-like G2P[4] RVA, with the genotype constellation G9-P[8]-I1-R1-C1-M1-A1-N1-T1-E2-H1 (G9P[8]-E2). Phylogenetic analysis indicated that N4006 had a common ancestor with Japanese G9P[8]-E2 rotavirus. Neutralizing epitope analysis showed that VP7, VP5*, and VP8* of N4006 had low homology with vaccine viruses of the same genotype and marked differences with vaccine viruses of other genotypes.
CONCLUSION
The RVA G9P[8] genotype with the G9-P[8]-I1-R1-C1-M1-A1-N1-T1-E2-H1 (G9P[8]-E2) constellation predominates in China and may originate from reassortment between Japanese G9P[8] with Japanese DS-1-like G2P[4] rotaviruses. The antigenic variation of N4006 with the vaccine virus necessitates an evaluation of the effect of the rotavirus vaccine on G9P[8]-E2 genotype rotavirus.
目的
分离一株中国流行的 G9P[8]型 A 群轮状病毒（RVA）（N4006），并对其基因组和进化特征进行研究，以期为新型轮状病毒疫苗的开发提供帮助。
方法
从腹泻样本中分离出的 RVA G9P[8]基因型在 MA104 细胞中传代。通过透射电镜、聚丙烯酰胺凝胶电泳和间接免疫荧光试验对病毒进行评估。采用 RT-PCR 和测序获得病毒的全基因组。利用 MEGA ver. 5.0.5 和 DNASTAR 软件对病毒的基因组和进化特征进行核酸序列分析。利用 BioEdit ver. 7.0.9.0 和 PyMOL ver. 2.5.2 分析 VP7 和 VP4（VP5和 VP8）的中和表位。
结果
RVA N4006（G9P[8]基因型）在 MA104 细胞中适应并获得高滴度（10 PFU/mL）。全基因组序列分析表明，N4006 是一株 Wa 样 G9P[8]RVA 和 DS-1 样 G2P[4]RVA 的 NSP4 基因的重组轮状病毒，基因型为 G9-P[8]-I1-R1-C1-M1-A1-N1-T1-E2-H1（G9P[8]-E2）。系统进化分析表明，N4006 与日本 G9P[8]-E2 轮状病毒具有共同的祖先。中和表位分析表明，N4006 的 VP7、VP5和 VP8与同基因型的疫苗病毒具有低同源性，与其他基因型的疫苗病毒存在明显差异。
结论
中国流行的 G9P[8]基因型以 G9-P[8]-I1-R1-C1-M1-A1-N1-T1-E2-H1（G9P[8]-E2）为特征，可能来源于日本 G9P[8]与日本 DS-1 样 G2P[4]轮状病毒之间的重组。N4006 与疫苗病毒的抗原变异需要评估轮状病毒疫苗对 G9P[8]-E2 基因型轮状病毒的效果。
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