EzMap: a simple pipeline for reproducible analysis of the human virome

Summary In solid-organ transplant recipients, a delicate balance between immunosuppression and immunocompetence must be achieved, which can be difficult to monitor in real-time. Shotgun sequencing of cell-free DNA (cfDNA) has been recently proposed as a new way to indirectly assess immune function in transplant recipients through analysis of the status of the human virome. To facilitate exploration of the utility of the human virome as an indicator of immune status, and to enable rapid, straightforward analyses by clinicians, we developed a fully automated computational pipeline, EzMap, for performing metagenomic analysis of the human virome. EzMap combines a number of tools to clean, filter, and subtract WGS reads by mapping to a reference human assembly. The relative abundance of each virus present is estimated using a maximum likelihood approach that accounts for genome size, and results are presented with interactive visualizations and taxonomy-based summaries that enable rapid insights. The pipeline is automated to run on both workstations and computing clusters for all steps. EzMap automates an otherwise tedious and time-consuming protocol and aims to facilitate rapid and reproducible insights from cfDNA. Availability and Implementation EzMap is freely available at https://github.com/dekoning-lab/ezmap. Contact jason.dekoning@ucalgary.ca. Supplementary information Supplementary data are available at Bioinformatics online.

[1]  Bartek Wilczynski,et al.  Biopython: freely available Python tools for computational molecular biology and bioinformatics , 2009, Bioinform..

[2]  R. Razonable,et al.  Human herpesvirus‐6 infections in kidney, liver, lung, and heart transplantation: review , 2012, Transplant international : official journal of the European Society for Organ Transplantation.

[3]  Li C. Xia,et al.  Accurate Genome Relative Abundance Estimation Based on Shotgun Metagenomic Reads , 2011, PloS one.

[4]  E. Delwart,et al.  The plasma virome of febrile adult Kenyans shows frequent parvovirus B19 infections and a novel arbovirus (Kadipiro virus). , 2016, The Journal of general virology.

[5]  P. Gunasekaran,et al.  Elevated Levels of Circulating DNA in Cardiovascular Disease Patients: Metagenomic Profiling of Microbiome in the Circulation , 2014, PloS one.

[6]  C. Desnues,et al.  Metagenomics and the Human Virome in Asymptomatic Individuals. , 2016, Annual review of microbiology.

[7]  F. Bushman,et al.  Viral Metagenomics Reveal Blooms of Anelloviruses in the Respiratory Tract of Lung Transplant Recipients , 2015, American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons.

[8]  Lance R. Martin,et al.  Noninvasive monitoring of infection and rejection after lung transplantation , 2015, Proceedings of the National Academy of Sciences.

[9]  Min Seong Kim,et al.  Single-stranded DNA library preparation uncovers the origin and diversity of ultrashort cell-free DNA in plasma , 2015, Scientific Reports.

[10]  N. Neff,et al.  Temporal Response of the Human Virome to Immunosuppression and Antiviral Therapy , 2013, Cell.