Find QIAseq xHYB AMR Markers (Human host)
The Find QIAseq xHYB AMR Markers (Human host) template workflow trims reads and detects antimicrobial resistance (AMR) markers.
It is suitable for analysis of samples from human hosts generated with the QIAseq xHYB AMR Panel.
To analyze non-human samples, you can create a copy of the workflow and edit it to fit your specific application, see https://resources.qiagenbioinformatics.com/manuals/clcgenomicsworkbench/current/index.php?manual=Template_workflows.html. Since the workflow element Map Reads to Human Control Genes is relevant for human data only, you should delete this.
Once the workflow copy is customized, you can install it to make it available from the Toolbox, see https://resources.qiagenbioinformatics.com/manuals/clcgenomicsworkbench/current/index.php?manual=Installing_workflow.html.
QIAGEN reference data set
The QIAseq xHYB AMR Panel reference data set is available from QIAGEN Sets Reference Data Library accessible via References () in the top Toolbar.
Launching the workflow
The Find QIAseq xHYB AMR Markers (Human host) workflow is at:
Toolbox | Template Workflows () | Microbial Workflows () | QIAseq Analysis () | Find QIAseq xHYB AMR Markers (Human host) ()
Launch the workflow and step through the wizard.
- Select the sequence list(s) containing the reads to analyze and click on Next.
- Select a reference data set or select "Use the default reference data" to configure the reference data elements individually in subsequent wizard steps (figure 2.77). Click on Next.
- Choose whether batch units should be defined based on organization of the input data, or by provided metadata (figure 2.78). For information on how to use metadata when running part of a workflow multiple times, see https://resources.qiagenbioinformatics.com/manuals/clcgenomicsworkbench/current/index.php?manual=Running_part_workflow_multiple_times.html.
- Next, you can review the batch units resulting from your selections above. Click on Next.
- Verify or select the reference marker database (figure 2.79). Click on Next.
- Verify or select control genes (figure 2.80). Click on Next.
- Specify the trim settings (figure 2.81) and click on Next.
- Finally, select a location to save outputs to and click on Finish.
Figure 2.77: Select reference data set.
Figure 2.78: Define batch units.
Figure 2.79: Select the reference marker database
Figure 2.80: Select control genes.
Workflow tools and outputs
The Find QIAseq xHYB AMR Markers (Human host) template workflow consists of the following tools. See figure 2.82 for a full overview of the workflow.
Figure 2.82: Layout of the QIAseq xHYB AMR Markers (Human host) workflow.
- QC for Sequencing Reads. Performs basic quality control of the sequencing reads. The output, which is included in a combined report, can be used to evaluate the quality of the sequencing reads. See https://resources.qiagenbioinformatics.com/manuals/clcgenomicsworkbench/current/index.php?manual=QC_Sequencing_Reads.html.
- Trim Reads. Removes adapter sequences and low quality nucleotides. The appropriate settings for the Trim Reads tool depends on the protocol used to generate the reads. See https://resources.qiagenbioinformatics.com/manuals/clcgenomicsworkbench/current/index.php?manual=Trim_Reads.html.
- Map Reads to Human Control Genes. Maps the host reads output from Taxonomic Profiling to the host taxonomic profiling index, to a reference of human control genes. See https://resources.qiagenbioinformatics.com/manuals/clcgenomicsworkbench/current/index.php?manual=Map_Reads_Reference.html. This serves as a QC step to verify mapping to the human control genes. For human samples, you expect to see mapping of reads to all human control genes.
- QC for Read Mapping. Performs quality control of the read mapping. See https://resources.qiagenbioinformatics.com/manuals/clcgenomicsworkbench/current/index.php?manual=QC_Read_Mapping.html.
- Find Resistance with ShortBRED. Detects and quantifies the presence of antimicrobial resistance marker genes of interest. See Find Resistance with ShortBRED.
- Merge Abundance Tables. Merges the sample-specific abundance tables to one combined abundance table. See Merge Abundance Tables.
The outputs provided by this workflow are:
- Resistance Table. The result table from Find Resistance with ShortBRED. The sample-specific output provides the abundance of each detected AMR marker. See Resistance abundance table.
- Read Mapping Human Control Genes. Sample-specific reads track containing the reads that mapped to the human control genes.
- Combined Report. Summary of the results from the individual tools for all the samples included in the workflow run.
- Merged Resistance Table. Provides the abundances of the detected AMR marker across samples. See Resistance abundance table.