北京某绿地蚊媒病毒的分离和鉴定

Isolation and Identification of Mosquito-borne Viruses at a Park in Beijing

为了解北京某绿地蚊媒病毒的分布情况,本试验使用诱蚊灯捕蚊采集到蚊虫标本19000余只,利用蚊子细胞C6/36培养进行病毒分离,利用高通量测序和逆转录聚合酶链式反应(RT-PCR)鉴定分离株;RT-PCR测定分离株病毒的序列,并与国内外相应病毒序列进行比对分析。结果显示,样本经病毒分离获得2株可引起明显细胞病变的分离株,分别命名为BJ2021/01和BJ2021/02。高通量测序和RT-PCR检测结果显示,BJ2021/01分离株含有版纳病毒(BAV)、忍者病毒(SHTV)、沙捞越病毒(SWKV)和库蚊源类芜菁黄花叶病毒科病毒(CuTLV),为4种病毒的混合物;BJ2021/02为BAV。基因序列分析结果显示,BAV分离株BJ2021/02的12个片段与国内外BAV分离株的核苷酸同源性为58.1%~98.7%,编码氨基酸的同源性为54.4%~98.9%。系统进化分析结果显示,BJ2021/02分离株属基因A型。本试验结果丰富了我国BAV的基因组序列,为开展北京市BAV的流行病学研究提供了参考,为虫媒病毒病的预防和控制提供了病原学依据。

In order to understand the distribution of mosquito-borne viruses in a park in Beijing,this study collected over 19000 mosquito specimens using mosquito trap lamps.Virus isolation was performed using mosquito cells C6/36,and the isolated strains were identified using high-throughput sequencing and reverse transcription-polymerase chain reaction(RT-PCR).The viral sequences of the isolated strains were determined by RT-PCR and compared with corresponding viral sequences from domestic and international sources.The results showed that two strains causing significant cytopathic effects were obtained through virus isolation,named BJ2021/01 and BJ2021/02.High-throughput sequencing and RT-PCR results indicated that the BJ2021/01 strain contained a mixture of Banna virus(BAV),Shinobi tetravirus(SHTV),Sarawak virus(SWKV),and Culex originated tymoviridae-like virus(CuTLV).The BJ2021/02 strain was identified as BAV.Sequence analysis revealed that the nucleotide homology of the 12 segments of the BAV BJ2021/02 strain with other domestic and international BAV strains ranged from 58.1%to 98.7%,and the amino acid homology ranged from 54.4%to 98.9%.Phylogenetic analysis indicated that the BJ2021/02 strain belonged to the BAV genotype A.These results enrich the genomic sequences of BAV in China,providing a reference for epidemiological research on BAV in Beijing and offering a pathogenic basis for the prevention and control of mosquito-borne viral diseases.