Phylogenetics has played a pivotal role in the genomic epidemiology of severe acute respiratory syndrome coronavirus 2, such as tracking the emergence and global spread of variants and scientific communication. However, the rapid accumulation of genomic data from around the world—with over two million genomes currently available in the Global Initiative on Sharing All Influenza Data database—is testing the limits of standard phylogenetic methods. Here, we describe a new approach to rapidly analyze and visualize large numbers of SARS-CoV-2 genomes. Using Python, genomes are filtered for problematic sites, incomplete coverage, and excessive divergence from a strict molecular clock. All differences from the reference genome, including indels, are extracted using minimap2 and compactly stored as a set of features for each genome. For each Pango lineage (https://cov-lineages.org), we collapse genomes with identical features into ‘variants’, generate 100 bootstrap samples of the feature set union to generate weights, and compute the symmetric differences between the weighted feature sets for every pair of variants. The resulting distance matrices are used to generate neighbor-joining trees in RapidNJ that are converted into a majority-rule consensus tree for each lineage. Branches with support values below 50 per cent or mean lengths below 0.5 differences are collapsed, and tip labels on affected branches are mapped to internal nodes as directly sampled ancestral variants. Currently, we process about 2 million genomes in approximately 9 h on 52 cores. The resulting trees are visualized using the JavaScript framework D3.js as ‘beadplots’, in which variants are represented by horizontal line segments, annotated with beads representing samples by collection date. Variants are linked by vertical edges to represent branches in the consensus tree. These visualizations are published at https://filogeneti.ca/CoVizu. All source code was released under an MIT license at https://github.com/PoonLab/covizu.

Significance Recombination is a major mechanism through which HIV type 1 (HIV-1) maintains genetic diversity and interferes with viral eradication efforts. There is growing evidence demonstrating a recombinant origin of primate lentiviruses including HIV-1 group M (HIV-1/M). Inferring the extent of recombination across the entire HIV-1/M genome is of great importance as it provides deeper insights into the origin, dynamics, and evolution of the global pandemic. Here we propose an alternative method that can reconstruct the extent of genome-wide recombination in HIV-1, uncover reticulate patterns, and serve as a framework for HIV-1 classification. Our method provides an alternative approach for understanding the roles of virus recombination in the early evolutionary history of zoonosis for other emerging viruses.

Phylogenetics has played a pivotal role in the genomic epidemiology of SARS-CoV-2, such as tracking the emergence and global spread of variants, and scientific communication. However, the rapid accumulation of genomic data from around the world - with over two million genomes currently available in the GISAID database - is testing the limits of standard phylogenetic methods. Here, we describe a new approach to rapidly analyze and visualize large numbers of SARS-CoV-2 genomes. Using Python, genomes are filtered for problematic sites, incomplete coverage, and excessive divergence from a strict molecular clock. All differences from the reference genome, including indels, are extracted using minimap2, and compactly stored as a set of features for each genome. For each Pango lineage (https://cov-lineages.org), we collapse genomes with identical features into 'variants', generate 100 bootstrap samples of the feature set union to generate weights, and compute the symmetric differences between the weighted feature sets for every pair of variants. The resulting distance matrices are used to generate neigihbor-joining trees in RapidNJ and converted into a majority-rule consensus tree for the lineage. Branches with support values below 50% or mean lengths below 0.5 differences are collapsed, and tip labels on affected branches are mapped to internal nodes as directly-sampled ancestral variants. Currently, we process about 1.6 million genomes in approximately nine hours on 34 cores. The resulting trees are visualized using the JavaScript framework D3.js as 'beadplots', in which variants are represented by horizontal line segments, annotated with beads representing samples by collection date. Variants are linked by vertical edges to represent branches in the consensus tree. These visualizations are published at https://filogeneti.ca/CoVizu. All source code was released under an MIT license at https://github.com/PoonLab/covizu.

Gene overlap occurs when two or more genes are encoded by the same nucleotides. This phenomenon is found in all taxonomic domains, but is particularly common in viruses, where it may provide a mechanism to increase the information content of compact genomes. The presence of overlapping reading frames (OvRFs) can skew estimates of selection based on the rates of non-synonymous and synonymous substitutions, since a substitution that is synonymous in one reading frame may be non-synonymous in another and vice versa. To understand the impact of OvRFs on molecular evolution, we implemented a versatile simulation model of nucleotide sequence evolution along a phylogeny with any distribution of open reading frames in linear or circular genomes. We use a custom data structure to track the substitution rates at every nucleotide site, which is determined by the stationary nucleotide frequencies, transition bias and the distribution of selection biases (dN/dS) in the respective reading frames. Our simulation model is implemented in the Python scripting language. All source codes are released under the GNU General Public License version 3 and are available at https://github.com/PoonLab/HexSE.

In previous work, we used a comprehensive sliding-window molecular clock analysis of near full length HIV-1 genomes to find that different regions of the virus genome yield significantly different estimates of the time to the most recent common ancestor. This finding, together with other evidence of the deep recombinant history of SIV and HIV, is not consistent with the standard model of the evolutionary history of HIV-1/M where non-recombinant subtypes are globally distributed through founder effects, followed by occasional inter-subtype recombination. We propose to re-examine the history of HIV-1/M using recombination detection methods that do not rely on predefined non-recombinant genomes. Here we describe an unsupervised non-parametric clustering approach to this problem by adapting a community detection method developed for the analysis of dynamic social networks. We compared our method to other reference-free recombination detection programs, namely GARD (HyPhy) and RDP (versions 4 and 5). We simulated recombination events by swapping randomly sampled branches in a time-scaled tree, which we reconstructed from subtype reference sequences in BEAST. Extant sequences were simulated for each tree using INDELible, and concatenated segments delimited by randomly sampled breakpoints from the resulting alignments to form the recombinant sequences. We show that our community detection method outperforms GARD, RDP4 and RDP5 in detecting recombinant breakpoints in simulated data, with a significantly lower mean error rate (Wilcoxon test, P < 0:05). Our method groups HIV-1 into 25 communities and detects evidence of inter-subtype recombination in pure subtype reference genomes obtained from Los Alamos HIV sequence database. For instance, we estimate that sub-subtypes A1 and A2 may contain large fragments from subtype C, while subtype C seems to contain fragments from subtype G. Our method provides a new reference-free framework for detecting recombination in viral genomes, and network communities may provide an alternative framework for HIV-1 classification.