Wiki¶

Data location:¶

The data was transferred from Athena medmicro):

/MedMicro/Clinton/CRE Pfizer Feb 2022/CRE study_1A_results_17022022 
/MedMicro/Clinton/CRE Pfizer Feb 2022/CRE study_1B_results_21022022

to Ilifu:

/scratch3/users/katiel/Clinton/CRE_study_August_2022/

Reference data:¶

Klebsiella pneumoniae – strain HS11286 (GenBank accession no. CP003200.1) (n=18);
Serratia marcescens – strain KS10 (GenBank accession no. CP027798.1) (n=3);
Escherichia coli – strain ATCC 25922 (GenBank accession no. CP009072.1) (n=1); and
Enterobacter cloacae – strain ATCC 13047 (GenBank accession no. NC_014121.1) (n=1).

/scratch3/users/katiel/Clinton/CRE_study_August_2022/ref_genomes

Objectives workflow:¶

QC:¶

11 sample failed QC phred scores before trimming and filtering; none failed after filtering and trimming. Filtering and trimming were executed as follows:

nextflow run kviljoen/fastq_QC --reads '/scratch3/users/katiel/Clinton/CRE_study_August_2022/raw/study_1A_B_combined/*_R{1,2}_001.fastq.gz' -profile ilifu

QC reports can be found in the 'files' tab

AMR profiling¶

The preference from Clinton is to do AMR profiling with the ResFinder DB. I'm getting errors there that I think relate to the header formatting though so in the interim have run with the ARG_annot DB that we used for previous projects as:

ARGannot¶

nextflow run kviljoen/uct-srst2 --reads '/scratch3/users/katiel/Clinton/CRE_study_August_2022/2022-09-19-fastq_QC/bbduk/*_{1,2}.fq' -profile ilifu --gene_db /cbio/users/katie/Nicol/Ps_aerug_srst2_MLST/ARGannot_r3.fasta --outdir /scratch3/users/katiel/Clinton/CRE_study_August_2022/srst2_ARGannot/coverage_80_run --min_gene_cov 80

Individual results files compiled as:

srst2 --prev_output *results.txt --output ARGannot_AMRs

CARD DB:¶

This database is the recommended by srst2 and has been formatted by them already. The DB was downloaded with:

wget https://github.com/katholt/srst2/blob/master/data/CARD_v3.0.8_SRST2.fasta?raw=true -O CARD_v3.0.8_SRST2.fasta

Pipeline execution as:

nextflow run kviljoen/uct-srst2 --reads '/scratch3/users/katiel/Clinton/CRE_study_August_2022/2022-09-19-fastq_QC/bbduk/*_{1,2}.fq' -profile ilifu --gene_db /scratch3/users/katiel/Clinton/CRE_study_August_2022/ref_files/CARD_v3.0.8_SRST2.fasta --outdir /scratch3/users/katiel/Clinton/CRE_study_August_2022/srst2_CARD/coverage_80_run --min_gene_cov 80

Individual results files compiled as:

srst2 --prev_output *results.txt --output CARD_AMRs