Recommendations for the introduction of metagenomic high-throughput sequencing in clinical virology, part I: Wet lab procedure.

Metagenomic high-throughput sequencing (mHTS) is a hypothesis-free, universal pathogen detection technique for determination of the DNA/RNA sequences in a variety of sample types and infectious syndromes. mHTS is still in its early stages of translating into clinical application. To support the development, implementation and standardization of mHTS procedures for virus diagnostics, the European Society for Clinical Virology (ESCV) Network on Next-Generation Sequencing (ENNGS) has been established. The aim of ENNGS is to bring together professionals involved in mHTS for viral diagnostics to share methodologies and experiences, and to develop application recommendations. This manuscript aims to provide practical recommendations for the wet lab procedures necessary for implementation of mHTS for virus diagnostics and to give recommendations for development and validation of laboratory methods, including mHTS quality assurance, control and quality assessment protocols.

[1]  A. Trkola,et al.  Two Years of Viral Metagenomics in a Tertiary Diagnostics Unit: Evaluation of the First 105 Cases , 2019, Genes.

[2]  Jose Manuel Martí,et al.  Recentrifuge: Robust comparative analysis and contamination removal for metagenomics , 2019, PLoS Comput. Biol..

[3]  Kristine M Wylie,et al.  Enhanced virome sequencing using targeted sequence capture , 2015, Genome research.

[4]  George G. Brownlee,et al.  Direct Evidence that the Poly(A) Tail of Influenza A Virus mRNA Is Synthesized by Reiterative Copying of a U Track in the Virion RNA Template , 1999, Journal of Virology.

[5]  W. Lipkin,et al.  Virome Capture Sequencing Enables Sensitive Viral Diagnosis and Comprehensive Virome Analysis , 2015, mBio.

[6]  O. Lund,et al.  Contaminating viral sequences in high-throughput sequencing viromics: a linkage study of 700 sequencing libraries. , 2019, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[7]  M. Feltkamp,et al.  Improved diagnosis of viral encephalitis in adult and pediatric hematological patients using viral metagenomics. , 2020, Journal of clinical virology : the official publication of the Pan American Society for Clinical Virology.

[8]  K. Metzner,et al.  Challenges and opportunities in estimating viral genetic diversity from next-generation sequencing data , 2012, Front. Microbio..

[9]  Rafi Ahmad,et al.  Nanopore-based DNA sequencing in clinical microbiology: preliminary assessment of basic requirements , 2018, bioRxiv.

[10]  Raju V. Misra,et al.  Development of a candidate reference material for adventitious virus detection in vaccine and biologicals manufacturing by deep sequencing , 2015, Vaccine.

[11]  Puthen V. Jithesh,et al.  Depletion of Human DNA in Spiked Clinical Specimens for Improvement of Sensitivity of Pathogen Detection by Next-Generation Sequencing , 2016, Journal of Clinical Microbiology.

[12]  Samuel S. Shepard,et al.  Direct RNA Sequencing of the Coding Complete Influenza A Virus Genome , 2018, Scientific Reports.

[13]  S. van Boheemen,et al.  Retrospective Validation of a Metagenomic Sequencing Protocol for Combined Detection of RNA and DNA Viruses Using Respiratory Samples from Pediatric Patients , 2019, The Journal of Molecular Diagnostics.

[14]  J. Mainardi,et al.  Untargeted next-generation sequencing-based first-line diagnosis of infection in immunocompromised adults: a multicentre, blinded, prospective study. , 2017, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[15]  J. Flanegan,et al.  Non-template functions of viral RNA in picornavirus replication. , 2011, Current opinion in virology.

[16]  M. Deijs,et al.  Viral Metagenomics on Cerebrospinal Fluid , 2019, Genes.

[17]  G. Reyes,et al.  Sequence-independent, single-primer amplification (SISPA) of complex DNA populations. , 1991, Molecular and cellular probes.

[18]  D. Kwiatkowski,et al.  Efficient Depletion of Host DNA Contamination in Malaria Clinical Sequencing , 2012, Journal of Clinical Microbiology.

[19]  Matthew Loose,et al.  Real-time selective sequencing using nanopore technology , 2016, Nature Methods.

[20]  Improved detection of artifactual viral minority variants in high-throughput sequencing data , 2015, Front. Microbiol..

[21]  P. Vende,et al.  Efficient Translation of Rotavirus mRNA Requires Simultaneous Interaction of NSP3 with the Eukaryotic Translation Initiation Factor eIF4G and the mRNA 3′ End , 2000, Journal of Virology.

[22]  Michael Huber,et al.  MinVar: A rapid and versatile tool for HIV-1 drug resistance genotyping by deep sequencing. , 2017, Journal of virological methods.

[23]  J. E. Muñoz-Medina,et al.  Metagenomic sequencing with spiked primer enrichment for viral diagnostics and genomic surveillance , 2020, Nature Microbiology.

[24]  Joseph L DeRisi,et al.  Actionable diagnosis of neuroleptospirosis by next-generation sequencing. , 2014, The New England journal of medicine.

[25]  Rémy Bruggmann,et al.  Limited Correlation of Shotgun Metagenomics Following Host Depletion and Routine Diagnostics for Viruses and Bacteria in Low Concentrated Surrogate and Clinical Samples , 2018, Front. Cell. Infect. Microbiol..

[26]  Brett J. Kennedy,et al.  Viral Pathogen Detection by Metagenomics and Pan-Viral Group Polymerase Chain Reaction in Children With Pneumonia Lacking Identifiable Etiology , 2017, The Journal of infectious diseases.

[27]  T. Mifflin Setting up a PCR laboratory. , 2007, CSH protocols.

[28]  I. Sidorov,et al.  Coronavirus discovery by metagenomic sequencing: a tool for pandemic preparedness , 2020, Journal of Clinical Virology.

[29]  Kai Zhao,et al.  A pneumonia outbreak associated with a new coronavirus of probable bat origin , 2020, Nature.

[30]  Colin R. Parrish,et al.  Presence and role of cytosine methylation in DNA viruses of animals , 2008, Nucleic acids research.

[31]  D. Deforce,et al.  Quantitative Bias in Illumina TruSeq and a Novel Post Amplification Barcoding Strategy for Multiplexed DNA and Small RNA Deep Sequencing , 2011, PloS one.

[32]  Marina N Nikiforova,et al.  Guidelines for Validation of Next-Generation Sequencing-Based Oncology Panels: A Joint Consensus Recommendation of the Association for Molecular Pathology and College of American Pathologists. , 2017, The Journal of molecular diagnostics : JMD.

[33]  T. Doan,et al.  Metagenomic Next-Generation Sequencing for Identification and Quantitation of Transplant-Related DNA Viruses , 2019, Journal of Clinical Microbiology.

[34]  Doug Stryke,et al.  Laboratory validation of a clinical metagenomic sequencing assay for pathogen detection in cerebrospinal fluid. , 2019, Genome research.

[35]  Doug Stryke,et al.  Clinical Metagenomic Sequencing for Diagnosis of Meningitis and Encephalitis. , 2019, The New England journal of medicine.

[36]  J. Keelan,et al.  Identification and removal of contaminating microbial DNA from PCR reagents: impact on low‐biomass microbiome analyses , 2018, Letters in applied microbiology.

[37]  D. Relman,et al.  Simple statistical identification and removal of contaminant sequences in marker-gene and metagenomics data , 2017, Microbiome.

[38]  Karsten Zengler,et al.  Improving saliva shotgun metagenomics by chemical host DNA depletion , 2018, Microbiome.

[39]  Niko Beerenwinkel,et al.  Error correction of next-generation sequencing data and reliable estimation of HIV quasispecies , 2010, Nucleic acids research.

[40]  Jacques Fellay,et al.  Viral Metagenomics in the Clinical Realm: Lessons Learned from a Swiss-Wide Ring Trial , 2019, Genes.

[41]  P. Simmonds,et al.  A European multicentre evaluation of detection and typing methods for human enteroviruses and parechoviruses using RNA transcripts , 2019, Journal of medical virology.

[42]  Marcos Parras-Moltó,et al.  Evaluation of bias induced by viral enrichment and random amplification protocols in metagenomic surveys of saliva DNA viruses , 2018, Microbiome.

[43]  V. Calvez,et al.  Fatal Encephalitis Caused by Cristoli Virus, an Emerging Orthobunyavirus, France , 2020, Emerging infectious diseases.

[44]  Martin S. Lindner,et al.  Analytical and clinical validation of a microbial cell-free DNA sequencing test for infectious disease , 2019, Nature Microbiology.

[45]  M. Gromeier,et al.  The hepatitis C virus 3′-untranslated region or a poly(A) tract promote efficient translation subsequent to the initiation phase , 2006, Nucleic acids research.

[46]  Robert Schlaberg,et al.  Validation of Metagenomic Next-Generation Sequencing Tests for Universal Pathogen Detection. , 2017, Archives of pathology & laboratory medicine.

[47]  Nagler Test UK Standards for Microbiology Investigations , 2014 .

[48]  C. Davis,et al.  Comparison of nucleic acid extraction methods for next-generation sequencing of avian influenza A virus from ferret respiratory samples. , 2019, Journal of virological methods.

[49]  John Hackett,et al.  The Perils of Pathogen Discovery: Origin of a Novel Parvovirus-Like Hybrid Genome Traced to Nucleic Acid Extraction Spin Columns , 2013, Journal of Virology.

[50]  David A. Matthews,et al.  Real-time, portable genome sequencing for Ebola surveillance , 2016, Nature.

[51]  K. Metzner,et al.  Next-Generation Sequencing of HIV-1 RNA Genomes: Determination of Error Rates and Minimizing Artificial Recombination , 2013, PloS one.

[52]  Angela C. M. Luyf,et al.  UvA-DARE ( Digital Academic Repository ) A Sensitive Assay for Virus Discovery in Respiratory Clinical Samples , 2011 .