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临床样本中宏基因组病毒检测方案。

Protocol for metagenomic virus detection in clinical specimens.

作者信息

Kohl Claudia, Brinkmann Annika, Dabrowski Piotr W, Radonić Aleksandar, Nitsche Andreas, Kurth Andreas

出版信息

Emerg Infect Dis. 2015 Jan;21(1):48-57. doi: 10.3201/eid2101.140766.

Abstract

Sixty percent of emerging viruses have a zoonotic origin, making transmission from animals a major threat to public health. Prompt identification and analysis of these pathogens are indispensable to taking action toward prevention and protection of the affected population. We quantifiably compared classical and modern approaches of virus purification and enrichment in theory and experiments. Eventually, we established an unbiased protocol for detection of known and novel emerging viruses from organ tissues (tissue-based universal virus detection for viral metagenomics [TUViD-VM]). The final TUViD-VM protocol was extensively validated by using real-time PCR and next-generation sequencing. We could increase the amount of detectable virus nucleic acids and improved the detection of viruses <75,000-fold compared with other tested approaches. This TUViD-VM protocol can be used in metagenomic and virome studies to increase the likelihood of detecting viruses from any biological source.

摘要

60%的新兴病毒源自动物,这使得动物传播成为公共卫生的一大威胁。及时识别和分析这些病原体对于采取行动预防和保护受影响人群至关重要。我们在理论和实验上对病毒纯化和富集的经典方法与现代方法进行了定量比较。最终,我们建立了一种从器官组织中检测已知和新型新兴病毒的无偏差方案(基于组织的病毒宏基因组学通用病毒检测 [TUViD-VM])。最终的TUViD-VM方案通过实时PCR和下一代测序进行了广泛验证。与其他测试方法相比,我们能够将可检测病毒核酸的量增加,并将病毒检测能力提高了<75,000倍。这种TUViD-VM方案可用于宏基因组学和病毒组研究,以提高从任何生物来源检测病毒的可能性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bbc9/4285256/7b4e8f474cac/14-0766-F2.jpg

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