山东省2021年蚊虫中Kadipiro病毒的全基因组序列特征

Complete Genome Characterization of Kadipiro Virus Isolated from Mosquitoes in Shandong Province,China in 2021

对2021年采集自山东省微山县蚊虫中的Kadipiro病毒(Kadipiro virus,KDV)进行分离鉴定和全基因组序列特征分析。将蚊虫标本研磨处理后接种细胞进行病毒分离,应用下一代测序技术对病毒分离物的全基因组序列进行测定,使用BioEdit 7.0.5.3软件进行相似性比较,使用Mega 11.0软件进行系统发生分析。成功分离到4株KDV,分别命名为SD21264、SD21397、SD21398和SD21399,均分离自三带喙库蚊。该4株KDV均能引起C6/36细胞产生明显的细胞病变,表现为细胞圆缩、间隙变大,但不能引起BHK21细胞病变。核苷酸相似性分析显示山东株中除了第7节段和第11节段之外,其他各节段互相之间的一致性大多>98.0%;与GenBank已有的4个KDV基因组序列相比,本研究分离株与我国2013年分离株(QTM27331)和2016年分离株(SDKL1625)的核苷酸一致性较高,其中片段1、3、6、9的一致性>98.0%;而与印尼株JKT-7075和丹麦株21164-6/M.dau/DK/2015的一致性较低,大多数片段在80%~90%之间。系统进化分析显示本研究分离株大部分节段与我国2016年分离株(SDKL1625)的进化关系最近,但7、11等部分节段有基因重排发生的证据。本研究结果丰富了Kadipiro病毒的全基因组信息。应加强对该病毒的监测,探索其与人类发热或脑炎等疾病的关联。

To isolate and identify the Kadipiro virus(KDV)from mosquitoes collected in Weishan County,Shandong Province,China in 2021,and analyze the characteristics of the whole genome sequence.Mosquito specimens were homogenized and inoculated on cultured cells for virus isolation.Next generation sequencing technology was used to obtain the whole genome sequence of virus isolates.Similarity comparison was conducted with BioEdit 7.0.5.3 software,and phylogenetic analysis were performed via Mega 11.0 software.Four KDV strains namely SD21264,SD21397,SD21398 and SD21399 were isolated from Culex tritaeniorhynchus.They caused obvious cytopathic effects on C6/36 cells,which characterized by cell shrinkage and gap enlargement,but could not cause pathological changes on BHK21 cell.Nucleotide similarity analysis showed that most of the other segments shared more than 98.0%identity with each other except the 7th and 11th segments of Shandong strain.Compared with four existing KDV genome sequences in GenBank,the isolates in this study had the highest nucleotide identity with the isolates in 2013(QTM27331)and 2016(SDKL1625)in China,among which the identity of segments 1,3,6 and 9 was more than 98.0%.However,it showed low identity with Indonesian strain JKT-7075 and Danish strain 21164-6/M.dau/DK/2015,and most fragments were between 80%and 90%similarity.The phylogenetic analysis showed that most segments of the isolate in this study showed the closest evolutionary relationship with the 2016 isolate in China(SDKL1625),but evidence of genetic rearrangement was observed for some segments such as 7 and 11.The results of this study enrich the whole genome information of Kadipiro virus.Surveillance should be strengthened to explore its relationship with fever,encephalitis or other human diseases.