Protocol for metagenomic virus detection in clinical specimens

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 c...

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Bibliographic Details
Published inEmerging infectious diseases Vol. 21; no. 1; pp. 48 - 57
Main Authors Kohl, Claudia, Brinkmann, Annika, Dabrowski, Piotr W, Radonić, Aleksandar, Nitsche, Andreas, Kurth, Andreas
Format Journal Article
LanguageEnglish
Published United States U.S. National Center for Infectious Diseases 01.01.2015
Centers for Disease Control and Prevention
Subjects
Animals
Callithrix
Chick Embryo
Diagnosis
Disease transmission
DNA, Viral - genetics
DNA, Viral - isolation & purification
emerging infectious diseases
Genes, Viral
High-Throughput Nucleotide Sequencing
Humans
infectious diseases
Medical research
Medicine, Experimental
Metagenome
Metagenomic Virus Detection in Clinical Specimens
metagenomics
Molecular Diagnostic Techniques
next generation sequencing
Real-Time Polymerase Chain Reaction
Signal-To-Noise Ratio
Virus diseases
Virus Diseases - diagnosis
virus purification
viruses
Zoonoses
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ISSN1080-6040
1080-6059
1080-6059
DOI10.3201/eid2101.140766

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Summary: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.
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ISSN:1080-6040
1080-6059
1080-6059
DOI:10.3201/eid2101.140766