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Scripts for ARIBA-based gene detection

Yu Wan

This repository consists of helper scripts for submitting ARIBA jobs or processing their outputs.


1. A Nextflow pipeline for ARIBA

This pipeline submits PBS (Portable Batch System) jobs of ARIBA to a high-performance computing (HPC) cluster with a control for queue sizes. Users may also opt to run ARIBA locally (without submitting jobs to the HPC system) using this pipeline. Two Nextflow scripts implement this pipeline:

  • ariba.nf: Main Nextflow script for the pipeline;
  • ariba.config: Configuration file of ariba.nf. By default, each PBS job requests four CPUs and 16 GB RAM from the system. The same request applies to local jobs as well.

Dependencies

  • Singularity-compatible Docker image of ARIBA
  • Nextflow v20.0 and above

Parameters

  • fastq: A glob for read files. It must be braced by a pair of quotation marks and no square brackets should be used. Default: "*_{1,2}.fastq.gz".
  • output_dir: Name and path for the parental directory of output files. Default: output.
  • db_dir: Name and path to access an existing ARIBA-compatible ResFinder database. Default: resfinder.
  • queue_size: Maximum number of concurrent PBS jobs submitted to the HPC system. Some computation facilities have restrictions for the queue size for fair use. Default: 10.

Example commands

# Install ARIBA's Singularity image through Docker
singularity pull docker://staphb/ariba  # Rename the Image file to ariba.sif

# Run the ARIBA Nextflow pipeline for gene detection
nextflow -Djava.io.tmpdir=$PWD run ariba.nf --fastq "$PWD/reads/*_{1,2}.fastq.gz" --db_dir $HOME/db/resfinder --output_dir output -c ariba.config -profile pbs --queue_size 15 -with-singularity $HOME/software/docker/ariba.sif

Outputs

Five subdirectories are created under the parental output directory by the pipeline:

  • /report: [sample]_report.tsv
  • /gene: [sample]_genes.fna, decompressed and renamed from ARIBA's output file assembled_genes.fa.gz.
  • /stats: [sample]_stats.tsv, renamed fromdebug.report.tsv.
  • /contig: [sample]_assemblies.fna.gz and [sample]_seqs.fna.gz, renamed from output files assemblies.fa.gz and assembled_seqs.fa.gz of each sample, respectively.
  • /log: [sample]_log_clusters.gz and [sample]_log_version.txt, renamed from output files log.clusters.gz and version_info.txt of each sample, respectively.

Users may refer to instructions for PAMmaker for a method converting this pipeline's outputs to an allelic presence-absence matrix.


2. Processing results of MLST analysis

compileMLST.py

Compile outputs (mlst_report.tsv, mlst_report.details.tsv, and report.tsv) from MLST analysis by ARIBA, assuming the names of ARIBA's output directories are genome names. Namely, the MLST reports should be generated with the command

ariba run db genome1_1.fastq.gz genome1_2.fastq.gz ./genome1

where an output directory shares the name of a genome.

Example command line

find ./* -maxdepth 1 -type d | python ~/ARIBA_toolkit/compileMLST.py

Outputs

  • STs.tsv: compiled from mlst_report.tsv files;
  • scores.tsv: compiled from report.tsv files, which include values of nucleotide identities;
  • hits.tsv: compiled from mlst_report.details.tsv, which contain values of reference coverage (%) and read depths of allele hits.

concMLSTalleles.py

Concatenate allele sequences of a given list of STs. It is useful for comparing STs at the nucleotide level and MLST-based phylogenetic construction. Currently this script does not support allele variants, namely, requires exact hits of query genomes to reference allele sequences.

Example command lines:

python concMLSTalleles.py -p st_profiles.tsv -s ./sequence_dir -e tfa > mlstSeqs.fna

# Append sequences to an extant FASTA file
python concMLSTalleles.py -p st_profiles_new.tsv -s ./sequence_dir -e tfa >> mlstSeqs.fna

This script assumes input ST profiles are offered in a tab-delimited file with columns of STs and names of MLST loci. (No other columns should be allowed) For example:

ST locus_1 locus_2 locus_3 locus_4 locus_5 locus_6 locus_7
1 11 21 31 41 51 61 71
2 12 21 31 41 51 61 71
... ... ... ... ... ... ... ...

The directory of MLST allele sequences is expected to be the same as that downloaded from the PubMLST database using ARIBA. (pubmlst_download). The '-s' parameter does not require an end forward slash for the path. By default, the delimiter for locus name (for instance, gene adk) and allele number in an allele name (for example, adk.1) is a period, although some databases use an underscore as the delimiter.

Dependencies: Python v3, packages pandas and biopython.