Tulloch, R.L.; Kok, J.; Carter, I.; Dwyer, D.E.; Eden, J.-S. An Amplicon-Based Approach for the Whole-Genome Sequencing of Human Metapneumovirus. Viruses2021, 13, 499.
Tulloch, R.L.; Kok, J.; Carter, I.; Dwyer, D.E.; Eden, J.-S. An Amplicon-Based Approach for the Whole-Genome Sequencing of Human Metapneumovirus. Viruses 2021, 13, 499.
Tulloch, R.L.; Kok, J.; Carter, I.; Dwyer, D.E.; Eden, J.-S. An Amplicon-Based Approach for the Whole-Genome Sequencing of Human Metapneumovirus. Viruses2021, 13, 499.
Tulloch, R.L.; Kok, J.; Carter, I.; Dwyer, D.E.; Eden, J.-S. An Amplicon-Based Approach for the Whole-Genome Sequencing of Human Metapneumovirus. Viruses 2021, 13, 499.
Abstract
Human metapneumovirus (HMPV) is an important cause of upper and lower respiratory tract disease in individuals of all ages. It is estimated that most individuals will be infected by HMPV by the age of 5 years old. Despite this burden of disease, there remains caveats in our knowledge of virus global genetic diversity due to a lack of HMPV sequencing, particularly at whole genome scale. The purpose of this study was to create a simple and robust approach for HMPV whole genome sequencing to be used for genomic epidemiological studies. To design our assay, all available HMPV full length genome sequences were downloaded from the NCBI GenBank database and used to design four primer sets to amplify long, overlapping amplicons spanning the viral genome and, importantly, specific to all known HMPV subtypes. These amplicons were then pooled and sequenced on an Illumina iSeq; however the approach is suitable to other common NGS platforms. We demonstrate the utility of this method using a representative subset of clinical samples and examine these sequences using a phylogenetic approach. Here we present an amplicon-based method for the whole genome sequencing of HMPV from clinical extracts that can be used to better inform genomic studies of HMPV epidemiology and evolution.
Keywords
human metapneumovirus; whole genome sequencing; genomic epidemiology
Subject
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
