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Efficient purification and concentration of viruses from a large body of high turbidity seawater

机译:从大量高浊度海水中有效纯化和浓缩病毒

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Graphical abstract Abstract Marine viruses are the most abundant entities in the ocean and play crucial roles in the marine ecological system. However, understanding of viral diversity on large scale depends on efficient and reliable viral purification and concentration techniques. Here, we report on developing an efficient method to purify and concentrate viruses from large body of high turbidity seawater. The developed method characterizes with high viral recovery efficiency, high concentration factor, high viral particle densities and high-throughput, and is reliable for viral concentration from high turbidity seawater. Recovered viral particles were used directly for subsequent analysis by epifluorescence microscopy, transmission electron microscopy and metagenomic sequencing. Three points are essential for this method: ? The sampled seawater (>150L) was initially divided into two parts, water fraction and settled matter fraction, after natural sedimentation. ? Both viruses in the water fraction concentrated by tangential flow filtration (TFF) and viruses isolated from the settled matter fraction were considered as the whole viral community in high turbidity seawater. ? The viral concentrates were re-concentrated by using centrifugal filter device in order to obtain high density of viral particles.
机译:图形摘要摘要海洋病毒是海洋中数量最多的实体,在海洋生态系统中起着至关重要的作用。但是,大规模了解病毒多样性取决于有效和可靠的病毒纯化和浓缩技术。在这里,我们报道了开发一种从大量高浊度海水中纯化和浓缩病毒的有效方法。该方法具有病毒回收效率高,浓缩因子高,病毒颗粒密度高,通量高的特点,对于高浊度海水中的病毒浓缩可靠。回收的病毒颗粒可直接用于落射荧光显微镜,透射电子显微镜和宏基因组测序进行后续分析。此方法必须要注意以下三点:经过自然沉淀后,首先将采样的海水(> 150L)分为水部分和沉降物质部分两部分。 ?通过切向流过滤(TFF)浓缩的水部分中的病毒和从沉降物部分中分离出的病毒都被视为高浊度海水中的整个病毒群落。 ?为了获得高密度的病毒颗粒,通过使用离心过滤装置将病毒浓缩物再浓缩。

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