基于病毒宏基因组技术侦测牡蛎体内病毒

Viral detection from oyster tissue using viral metagenomics technology

  • 摘要: 建立了牡蛎(Crassostrea spp.)病毒宏基因组研究方法,应用差速离心和蔗糖垫超速离心提取病毒样颗粒,核酸酶处理宿主游离核酸后提取病毒总核酸,经反转录、双链合成和核酸扩增获得足量DNA。高通量测序共获得1 661 809 000个碱基,包括6 647 236个读长(reads)信息,拼接后得到50 842个重叠群(contig),序列总长度达到14 239 389 bp。同时从reads质控图等多方面对测序数据进行分析与评估,显示测序数据质量良好。数据与病毒库进行比对,发现病毒序列来自于小核糖核酸病毒科(Picornaviridae)和疱疹病毒科(Herpesviridae)2个科,类单纯疱疹病毒(Simplexvirus)、心病毒(Cardiovirus)和肠道病毒(Enterovirus)3个病毒属。

     

    Abstract: In order to work out a viral metagenomic research method of oyster (Crassostrea spp.), we applied differential centrifugation and ultracentrifugation with sucrose gradient to obtain the virus-like particle in crude extract. After further nuclease treatment of host nucleic acid, reverse transcription and double-stranded amplification, we used Hiseq2500 platform to produce a viral genome dataset from the sample. Results show that a total of 1 661 809 000 nucleotides, including 6 647 236 reads information were obtained and further overlapped into 50 842 contigs whose total sequence length reached 14 239 389 bp. Besides, we observed good quality of reads by the control chart and so on. We scanned the GenBank virus database with data, which indicates that the known virus sequences belonged to two families (Picornaviridae, Herpesviridae) and three virus genus (Simplexvirus, Cardiovirus, Enterovirus).

     

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