tbvariantfilter

This tool offers multiple options for filtering variants (in VCF files, relative to M. tuberculosis H37Rv coordinates).

It currently has 5 main modes:

1. Filter by region. Mask out variants in certain regions. Region lists available as:
   1. farhat_rlc and farhat_rlc_lowmap: Refined Low Confidence (RLC) and RLC plus low mappability regions from Marin et al
   2. pe_ppe: PE/PPE genes from Fishbein et al 2015
   3. tbprofiler: TBProfiler list of antibiotic resistant genes
   4. mtbseq: MTBseq list of antibiotic resistant genes
   5. uvp: UVP list of repetitive loci in M. tuberculosis genome
2. Filter by proximity to indels. Masks out variants within a certain distance (by default 5 bases) of an insertion or deletion site.
3. Filter by percentage of alternate allele bases. Mask out variants with less than a minimum percentage (by default 90%) alternative alleles.
4. Filter by depth of reads at a variant site. Masks out variants with less than a minimum depth of coverage (default 30) at the site
5. Filter all non-SNV variants. Masks out variants that are not single nucleotide variants.

Filtering by (SAM/BAM) mapping quality was omitted because these filters are performed by the upstream workflow we (SANBI) currently use.

When used together the effects of the filters are added (i.e. a variant is masked out if it is masked by any of the filters).

Installation

The software is available via bioconda and can be installed with:

conda install tb_variant_filter

Usage

``` usage: tbvariantfilter [-h] [--regionfilter REGIONFILTER] [--closetoindelfilter] [--indelwindowsize INDELWINDOWSIZE] [--minpercentagealt_filter] [--minpercentagealt MINPERCENTAGEALT] [--mindepthfilter] [--mindepth MINDEPTH] [--snvonlyfilter] inputfile [output_file]

Filter variants from a VCF file (relative to M. tuberculosis H37Rv)

positional arguments: inputfile VCF input file (relative to H37Rv) outputfile Output file (VCF format)

optional arguments: -h, --help show this help message and exit --regionfilter REGIONFILTER, -R REGIONFILTER --closetoindelfilter, -I Mask out single nucleotide variants that are too close to indels --indelwindowsize INDELWINDOWSIZE Window around indel to mask out (mask this number of bases upstream/downstream from the indel. Requires -I option to selected) --minpercentagealtfilter, -P Mask out variants with less than a given percentage variant allele at this site --minpercentagealt MINPERCENTAGEALT Variants with less than this percentage variants at a site will be masked out --mindepthfilter, -D Mask out variants with less than a given depth of reads --mindepth MINDEPTH Variants at sites with less than this depth of reads will be masked out --snvonly_filter Mask out variants that are not SNVs ```

Note that there are no filters by default. Each filter to be used needs to be explicitly mentioned.

To export a region (from the list of possible region masks) in BED format, use the tb_region_list_to_bed command:

``` usage: tbregionlisttobed [-h] [--chromosomename CHROMOSOMENAME] {farhatrlc,farhatrlclowmap,mtbseq,peppe,tbprofiler,uvp} [output_file]

Output region filter in BED format

positional arguments: {farhatrlc,farhatrlclowmap,mtbseq,peppe,tbprofiler,uvp} Name of region list output_file File to write output to

optional arguments: -h, --help show this help message and exit --chromosomename CHROMOSOMENAME Chromosome name to use in BED ```

Testing and development environment

The repository contains a file, test_environment.yml, for creating a conda environment for testing and development. Tests can be run with pytest and tox, where tox also uses conda to create the testing environment.

For some tests, locus locations are looked up using the COMBAT-TB NeoDB. This requires an environment variable, COMBATTB_BOLT_URL. If this is not set, tests requiring this lookup are skipped. The default in tox.ini uses the SANBI hosted NeoDB instance.

Licensing and distribution

This code free software and is licensed under the terms specified in COPYING, i.e under the terms of the GNU General Public License version 3.