广西中越边境口岸地区1株Nam Dinh病毒的分离及鉴定  

First isolation and identification of Nam dinh virus at the area of China-Vietnamese border port in Guangxi

作　　者：梁亮[1] 陈世毅[1] 李又云 韦一知[1] 邓秋云[1] 邓丽丽[1] 韦敬航[1] 马宇燕[1] 秦月[1] 刘巍[1] LIANG Liang;CHEN Shi-yi;LI You-yun;WEI Yi-zhi;DENG Qiu-yun;DENG Li-li;WEI Jing-hang;MA Yu-yan;QIN Yue;LIU Wei(Guangxi Center for Disease Control and Prevention,Nanning,Guangxi;Ningming Center for Disease Control and Prevention)

机构地区：[1]广西壮族自治区疾病预防控制中心,广西南宁530028 [2]宁明县疾病预防控制中心

出　　处：《中国病原生物学杂志》2022年第10期1121-1124,共4页Journal of Pathogen Biology

基　　金：广西壮族自治区卫生和计划生育委员会自筹经费课题(No.Z20180986)。

摘　　要：目的分离鉴定广西中越边境口岸地区来源的1株NDiV,并对其进行同源性比较和进化分析。方法将蚊虫标本研磨液接种C6/36和BHK21细胞,提取阳性分离物的病毒RNA,合成cDNA后进行二代测序,获得毒株的全基因组序列,利用Editseq软件识别该序列的ORF和非编码区,采用Bioedit 7.0软件进行同源性比较,采用MegaⅩ软件分别构建全基因组、RDRP和S基因的系统进化树。结果接种1批次三带喙库蚊标本的C6/36细胞出现明显的病变,病毒命名为Ad2019035sd,二代测序显示该毒株的NDiV覆盖率为99.80%;比较Ad2019035sd与Mesoniviridae科参考株全基因组核苷酸同源性在57.9%~98.9%之间,主要蛋白的编码基因核苷酸和氨基酸同源性分别在56.7%~100%和40.0%~99.5%之间,其中与深圳毒株SZ11706Z的同源性最高。全基因组、RDRP和S基因系统进化树均显示毒株Ad2019035sd与两株NDiV毒株在同一进化分支。结论毒株Ad2019035sd为NDiV,与深圳毒株SZ11706Z高度同源,为广西地区NDiV感染情况调查及相关研究提供了基础资料。Objective To identify one strain of NDiV isolated from the area of China-Vietnamese border port in Guangxi,and perform homology comparison and evolutionary analysis with other strains.Methods The mosquito grinding supernatant was used to infect C6/36 and BHK-21 cells respectively,the viral RNA of positive isolate was extracted,and its cDNA was synthesized for next-generation sequencing to obtain the whole genome sequence of the strain.The ORFs and the non-coding regions of the sequence were identified using Editseq,nucleotide and amino acid sequences of the strain were compared to those of other strains using Bioedit 7.0,and phylogenetic trees for complete genome,RDRP and S gene were constructed using Mega X.Results Obvious cytopathic effect was observed in C6/36 cell inoculated with one batch of Culex tritaeniorhynchus,and this strain was named Ad2019035 sd.The result of next-generation sequencing showed that NDiV coverage of this strain was 99.80%,in comparison of the nucleotide and amino acid homology with other reference strains of Mesoniviridae,the genomic nucleotide similarity ranged from 57.9%to 98.9%,and the nucleotide and amino acid similarities of important NDiV proteins ranged from 56.7%to 100%and 40.0%to 99.5%,respectively,and the Ad2019035 sd strain had the highest similarity with the Shenzhen strain(SZ11706 Z);furthermore,phylogenetic trees based on complete genome,RDRP and S gene also indicated that the Ad2019035 sd strain and two NDiV strains were on the same evolutionary branch.Conclusion The Ad2019035 sd strain belongs to NDiV and has high homology with the Shenzhen strain(SZ11706 Z),which could serve as an experimental material for further research on NDiV infection in Guangxi.

关 键 词：Nam Dinh病毒 中越边境口岸 全基因组 系统进化树 

分 类 号：R373.1[医药卫生—病原生物学]