食品科学 ›› 2018, Vol. 39 ›› Issue (2): 287-292.doi: 10.7506/spkx1002-6630-201802045

• 安全检测 • 上一篇    下一篇

应用Illumina Miseq测序分析饮用水源水中病毒多样性

葛英亮1,2,于水利2,3,*,时文歆2,*   

  1. (1.哈尔滨学院食品工程学院,黑龙江?哈尔滨 150080;2.哈尔滨工业大学市政环境工程学院,黑龙江?哈尔滨 150090;3.同济大学环境科学与工程学院,上海 200092)
  • 出版日期:2018-01-25 发布日期:2018-01-05
  • 作者简介:葛英亮,于水利,时文歆
  • 基金资助:
    “十二五”国家科技重大专项(2012ZX07403-001)

Analysis of Virus Diversity in Drinking Source Water by Using Illumina MiSeq Sequencing Technology

GE Yingliang1,2, YU Shuili2,3,*, SHI Wenxin2,*   

  1. (1. School of Food Engineering, Harbin University, Harbin 150080, China;2. School of Municipal and Environmental Engineering, Harbin Institute of Technology, Harbin 150090, China;3. College of Environmental Science and Engineering, Tongji University, Shanghai 200092, China)
  • Online:2018-01-25 Published:2018-01-05

摘要: 以东太湖水源水为研究对象,应用梯度-串联-循环-切向流超滤技术对水体中病毒进行分离浓缩,使用非序列依赖性单引物扩增技术扩增源水中病毒基因组,采用Illumina Miseq进行测序,与NCBI基因数据库进行比对,质控后获得1?190?914?928?bp基因数据,可组装成5?554?条scaffolds序列,获知病毒基因组功能,经比对可注释到尾噬菌体目(Caudovirales)、疱疹病毒目(Herpesvirales)、线状病毒目(Ligamenvirales),在科的水平上,注释到40?个科病毒的同源序列,其中含量较高的病毒科为Microviridae占27.590?1%,Siphoviridae占23.010?7%,Phycodnaviridae占5.322?2%,Retroviridae占1.691?2%,Mimiviridae占1.960?8%,其中无法注释(norank)占21.572?8%,102?112?条reads在科水平上norank,研究可以高通量的获得水源水中病毒多样性信息,并为水体中病毒的检测提供理论参考依据。

关键词: Illumina Miseq测序, 饮用水源水, 病毒, 多样性

Abstract: The viruses in drinking source water from east Tai Lake were separated and concentrated by gradient-series connection-circulation-tangential flow ultrafiltration (GSC-TFF). The viral genome was amplified by sequence independent single primer amplification (SISPA), sequenced by Illumina Miseq, and compared with the NCBI gene database using the basic local alignment search tool (BLAST). After quality control, 1 190 914 928 bp gene data were obtained, which could be assembled into 5 554 scaffold sequences. Viral genome function was found and annotated to Caudovirales, Herpesvirales and Ligamenvirales at the family level. A total of 40 families of homologous sequences were annotated, Microviridae (27.590 1%), Siphoviridae (23.010 7%), Phycodnaviridae (5.322 2%), Retroviridae (1.691 2%), Mimiviridae (1.960 8%) being the dominant ones. A total of 102 112 reads (21.572 8%) were identified as no rank at the family level. The approach proposed in this study can allow high throughput analysis of virus diversity in source water, paving the foundation for?detecting virus in water.

Key words: Illumina Miseq sequencing, drinking source water, virus, diversity

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