Workflows¶
Current available workflows are implemented in the folder workflows. Each workflow will depend on rules, stored in the folder of the same name, and can also depend on other workflows. rules are sorted with respect to their general function in different folders.
Core genome determination¶
Core genomes can be calculated by three different means.
Ridom¶
cgMLST scheme from ridom can be extracted directly for theses species
| Species | Taxonomy ID | Ridom ID | Reference genome assembly ID |
|---|---|---|---|
| Staphylococcus aureus | 1280 | 141106 | 33148 |
| Mycobacterium tuberculosis | 1773 | 741110 | 538048 |
| Listeria monocytogenes | 1639 | 690488 | 264498 |
| Escherichia coli | 562 | 5064703 | 79781 |
| Klebsiella pneumoniae | 573 | 2187931 | 31388 |
| Enterococcus faecium | 1352 | 991893 | 526908 |
| Acinetobacter baumannii | 470 | 3956907 | 39528 |
| Legionella pneumophila | 446 | 1025099 | 30068 |
A bed file is constructed from the locus target file, constructing coordinates from the start and length columns of the csv file file available on the ridom website.
Example target file:
snakemake --snakefile $pipeline_folder/workflows/core_genome/make_ridom.rules core_genomes/Staphylococcus_aureus/ridom/33148.bed will create the BED file defining the core genomic regions in the genome of the assembly ID 33148 (Staphylococcus aureus COL)
Enterobase¶
cgMLST scheme from enterobase is extracted for Salmonella enterica:
| Species | Taxonomy ID | Enterobase ID | Reference genome assembly ID | Scheme |
|---|---|---|---|---|
| Salmonella enterica | 28901 | SALwgMLST | 359488 | cgMLSTv1 |
A bed file for the reference genome 359488, based on the locus tag present in this genome is constructed. For instance, over the 3002 locus of the Salmonella cgMLSTv1, 69 come from a different genome than the reference 359488.
snakemake --snakefile $pipeline_folder/workflows/core_genome/make_enterobase.rules core_genomes/Salmonella_enterica/ridom/359488.bed will create the BED file defining the core genomic regions in the genome of the assembly ID 359488 (Salmonella enterica subsp. enterica serovar Typhimurium str. D23580)
ParSNP¶
For species unavailable on either resource, core genome can be calculated using parsnp and the complete genomes of the species available on RefSeq. As ParSNP is not available on bioconda, the binary must be downloaded from the ParSNP website and placed in your $PATH.
Assembly and quality¶
Aggregates rules for assembling genomes and performing various quality control checks. Required parameters:
cov_cutoff: contigs whose coverage is below this cutoff will be excluded from the final assemblyadapter_file_name: look for the adaptor for this library preparation kit (possible values)adapter_removal_param1,adapter_removal_param2,adapter_removal_param3: parameters for adapter trimming (reference)minimum_quality_base: leading and trailing bases below this quality will be removedminimum_read_length: reads shorter than this threshold after trimming will be discarded (be careful when using reads from SRA!)
Deliverables:
quality/multiqc/self_genome/multiqc_report.html: quality control report based on the results of fastqc, trimmomatic, qualimap, quast and prokka for every samplesamples/{sample_name}/annotation/: folder containing all annotation files from theprokkasoftware
Resistance¶
Depends on the Assembly and quality workflow.
Required parameters:
resistance_prediction_softwares: list of software for genetic resistance assessment. Possible values:mykrobeandrgi.
Deliverables:
samples/{sample_name}/annotation/resistance/rgi.tsv: results files for RGIsamples/{sample_name}/annotation/resistance/mykrobe.tsv: results file for mykrobe
Virulence¶
Depends on the Assembly and quality workflow.
Required parameters:
virulence_factors: file with list of uniprot accession of virulence factors. An example is available in the folderdata/staph/db/
Deliverables:
virulence_summary.xlsx: summary of virulence proteins found in every samples.
Epidemiology¶
Depends on the Assembly and quality workflow (for determining the Sequence Types).
Required parameters:
minimum_coverage_for_calling: minimum of coverage for considering a genomic position when counting differences between samples. Any position (SNP or non-SNP when compared to the reference) having a lower coverage will be maskedminimum_alternate_fraction_for_calling: minimum ratio of observations favouring a SNP over observations not favouring a SNP. Any SNPs not meeting this criteria will also be masked
Deliverables:
typing/{snp_caller}/core_{ridom or enterobase}/{reference_genome}/bwa/distance_snp_mst_no_st.svg: Minimum spanning tree of the distance in snps between every sample over the core genome as defined by ridom or enterobase. Available species and values for reference genomes are listed in the files indata/core_genome_dbs/. If the species under consideration has a multiple locus sequence type available,typing/{snp_caller}/core_{ridom or enterobase}/{reference_genome}/bwa/distance_snp_mst_with_st.svgcan be generated with the ST of each sample.phylogeny/{snp_caller}/core_{ridom or enterobase}/{reference_genome}/bwa/phylogeny_no_st.svg: A phylogeny based on the alignments of the core SNPs, using RAxML. Available species and values for reference genomes are listed in the files indata/core_genome_dbs/. If the species under consideration has a multiple locus sequence type available,phylogeny/{snp_caller}/core_{ridom or enterobase}/{reference_genome}/bwa/phylogeny_with_st.svgcan be generated with the ST of each sample.quality/multiqc/mapping_to_{reference_genome}/multiqc_report.html: multiqc report of qualimap, fastqc and trimmomatic of every samples when mapping against the reference. Check for quality control.