Pre-built MLST databases
Pre-built MLST databases are provided here and updated weekly: https://github.com/jordanlab/STing_datasets
We recommend building a new database each time STing is run, or daily if under continual use, to ensure the most up to date definitions as possible
Typing
Preparing directory:
mkdir STing_demo
cd STing_demo
Downloading the Neisseria spp. MLST database from PubMLST.org and build a STing index from it:
db_util.py fetch --query "Neisseria spp." --out_dir my_dbs --build_index
Downloading a WGS sample to analyze:
wget ftp://ftp.sra.ebi.ac.uk/vol1/fastq/ERR026/ERR026529/ERR026529_1.fastq.gz ftp://ftp.sra.ebi.ac.uk/vol1/fastq/ERR026/ERR026529/ERR026529_2.fastq.gz
Run STing typer:
typer -x my_dbs/neisseria_spp/db/index -1 ERR026529_1.fastq.gz -2 ERR026529_2.fastq.gz -s ERR026529 -c -a -d -t ERR026529.depth.tsv -y -o ERR026529.results.tsv --sensitive
Gene detection
Prbuilt CARD and VFDB databases are available on the STing Datasets repository: https://github.com/jordanlab/STing_datasets
Both pre-built databases and the one described below are for use with detector
.
Preparing directory:
mkdir -p STing_demo/my_dbs/card
cd STing_demo/my_dbs/card
Preparing the database:
Downloading the antimicrobial resistance gene database from CARD
wget https://card.mcmaster.ca/latest/data -O card.tgz
tar xvf card.tgz
Changing IUPAC extended DNA characters to N on the CARD nucleotide file:
sed -e '/^[^>]/s/[^ATGCatgc]/N/g' nucleotide_fasta_protein_homolog_model.fasta > card.fasta
Creating STing db config file:
echo -e "[loci]\ncard\tcard.fasta\n" > config.txt
The new config file should look like this:
[loci]
card card.fasta
Creating the STing database index:
indexer -c config.txt -p db/index -m GDETECT
Downloading a WGS sample to analyze:
wget ftp://ftp.sra.ebi.ac.uk/vol1/fastq/ERR026/ERR026529/ERR026529_1.fastq.gz ftp://ftp.sra.ebi.ac.uk/vol1/fastq/ERR026/ERR026529/ERR026529_2.fastq.gz
Run STing detector:
cd ../../
typer -x my_dbs/neisseria_spp/db/index -1 ERR026529_1.fastq.gz -2 ERR026529_2.fastq.gz -s ERR026529 -c -a -d -t ERR026529.depth.tsv -y -o ERR026529.results.tsv --sensitive