The metagenomics classifier uses a k-mer based classification algorithm to classify each query sequence (usually a read) against a collection of reference sequences. There are two logical steps to this process: 1) reference sequences are indexed into a searchable database 2) reference sequence database is searched using query sequences and query sequences are classified to taxid(s) associated with the reference sequences. This guide explains how to run query sequences against a pre-existing reference sequence database (As of DRAGEN 4.3+, users can build their own custom reference sequence database).
| Option | Description |
|---|---|
Applies to: --kmer-classifier-input-read-file, --kmer-classifier-multiple-inputs
If the analysis is for a single FASTA/FASTQ read file, then that filename is input to --kmer-classifier-input-read-file and --kmer-classifier-multiple-inputs=false. However, many read files can be submitted to the Kmer Classifier at one time, minimizing the load time for a large reference sequence database. In this case, the input file must be a .tsv (tab-separated) file with two columns (optionally 3 columns). The first column is a unique ID, the second column is the path to the read file, and the optional third column is the path to the second read file in the case of paired-end reads. The ID is used to distinguish the output files. There is no header line. This .tsv file is the input file to --kmer-classifier-input-read-file and --kmer-classifier-multiple-inputs=true.
Applies to: --kmer-classifier-db-file, --kmer-classifier-db-to-taxid-json, --kmer-classifier-load-db-ram
A file of reference sequences (the "database") can be quite large. If the database file is stored on a normal file system, it is recommended that you set --kmer-classifier-load-db-ram=true. This will tell the Kmer Classifier to load the database file into memory for faster analysis. It is also allowable to store the database file on a RAM disk, which reduces load time over many Kmer Classifier runs. In this case, it is recommended to set --kmer-classifier-load-db-ram=false.
Applies to: --kmer-classifier-db-to-taxid-json
This input file is downloaded alongside the reference sequence database. It associates a taxid internal to the classifier database to an external source, like the NCBI taxonomy. This JSON file is a dictionary where the keys are internal taxids, and is mapped to an external taxid, name, and rank. Example:
The internal taxids are used in the output files. This JSON file can be used to map the results to taxids from the NCBI taxonomy.
The genome database includes NCBI RefSeq genomes for human, bacteria, archaea, viruses, and fungi. The December 3 2023 NCBI taxonomy was used to build the database, and the sequences were collected in December 2023.
To download the reference index file and the taxid mapping JSON:
This database includes the contents of the Genome database and all of the NCBI nucleotide (nt) database. The sequences from the NCBI nucleotide database were collected in July 2023, and the December 3 2023 NCBI taxonomy was used to build the database. Two versions of this database are available for download: One that requires a machine with >= 550GB RAM, and a compressed version that trades approximately 5-10% accuracy for a smaller RAM footprint and requires a machine with >= 225GB RAM.
To download the reference index file and the taxid mapping JSON:
To download the compressed reference index file and the taxid mapping JSON:
This database includes all protein sequences of the UniRef90 database. The sequences were collected in March 2024 and the March 28 2024 NCBI taxonomy was used to build the database.
To download the reference index file and the taxid mapping JSON:
This database includes full length bacterial 16S sequences from the NCBI. The sequences were collected in April 2024 and the March 28 2024 NCBI taxonomy was used to build the database.
To download the reference index file and the taxid mapping JSON:
There are two output files, one organized around the reads, and the other organized around the taxids.
Applies to: --kmer-classifier-output-taxid-seq, --kmer-classifier-output-read-seq The main output file is a .tsv file with the extension .read_classifications.tsv. It has no header line, has tab-separated columns, and can vary in the number of columns depending on command line options. It details the results for each read.
The second output file is a .tsv file with the extension .classifier.taxid_kmer_counts.tsv. It has a header line and has tab-separated columns. It summarizes the results for each taxid.
The metagenomics classifier uses a k-mer based classification algorithm to classify each query sequence (usually a read) against a collection of reference sequences. There are two logical steps to this process: 1) reference sequences are indexed into a searchable database 2) reference sequence database is searched using query sequences and query sequences are classified to taxid(s) associated with the reference sequences. This guide explains how to generate your own indexed, searchable database of reference sequences to be used by the k-mer classifier.
| Option | Description |
|---|
K-mer/G-mer length considerations:
G-mer length refers to the size of the window from which to pick a minimizer. The larger the window, the fewer minimizers will be chosen overall, resulting in a smaller database. However, this can cause a loss of sensitivity since fewer k-mers overall will be saved.
K-mer length refers to the size of the minimizer to be saved in a window size specified by the G-mer length. In general, larger k-mers result in greater specificity while shorter k-mers result in greater sensitivity. However, this general statement can be proven wrong by the specifics of an application and we recommend trying a few different g-mer and k-mer lengths to determine what works best for a given sequence type and application.
As a general rule, we recommend starting with a g-mer length of 35 and k-mer length of 31.
Pre-built Explify Reference Database k-mer/g-mer length settings for reference:
Very large collection of NCBI Refseq genomes and the entirety of the NCBI nucleotide (nt) database (more than 2 Terabases of sequence): G-mer length of 41 and k-mer length of 31. Compressed version built with g-mer length of 64 and k-mer length of 31. This results in a database with less than half the storage requirements.
Subset of reference genomes from Refseq with a focus on viral detection: G-mer length of 35 and k-mer length of 31
Collection of 16S sequences for bacterial identification / profiling: G-mer length of 31 and k-mer length of 31
Uniref90 protein sequences: G-mer length of 15, k-mer length of 12 (these are k-mers of amino acids, not nucleotides)
Three types of databases can be built with this tool:
Binner: each k-mer is assigned to a category/bin.
Must use --kmer-class-db-builder-num-categories.
Do not use --kmer-class-db-builder-tax-tree-file, --kmer-class-db-builder-save-weights, or --kmer-class-db-builder-kmer-cutoff.
Classifier: each k-mer is assigned to one taxid.
Must define a taxonomic tree with --kmer-class-db-builder-tax-tree-file.
Do not use --kmer-class-db-builder-num-categories, --kmer-class-db-builder-save-weights, or --kmer-class-db-builder-kmer-cutoff.
Classifier with weights: each k-mer is assigned to one or more taxids; associated weights are also stored (frequency of k-mer's association with a taxid). Uses much more memory, but is more accurate.
Must use --kmer-class-db-builder-save-weights and define a taxonomic tree with --kmer-class-db-builder-tax-tree-file.
Can use --kmer-class-db-builder-kmer-cutoff.
Do not use --kmer-class-db-builder-num-categories.
| Column | Description | Data Type |
|---|---|---|
| Header | Description | Data Type |
|---|---|---|
Required Inputs
--enable-kmer-classifier
Enables the Kmer Classifier. (Default=false).
--output-file-prefix
Prefix for all output files.
--output-directory
Directory for all output files.
--kmer-classifier-input-read-file
Input sequence file (zipped or unzipped) to the Kmer Classifier.
--kmer-classifier-db-file
Database of sequences to classify against.
Optional Inputs
--intermediate-results-dir
Area for temporary files. Size must be greater than size of all FASTQ files multiplied by 2.
--kmer-classifier-load-db-ram
Load the database onto RAM. Do not use if database is on ramdisk. (Default=false).
--kmer-classifier-multiple-inputs
Set to true to run with multiple inputs. The input read file is now a .tsv file that has three columns: Sample ID, Read1 file, (optional) Read 2 file. (Default=false).
--kmer-classifier-min-window
The minimum number of consecutive kmers to classify assignment at taxid. (Default=1).
--kmer-classifier-output-read-seq
Option to enable read sequence column in the output file. (Default=false).
--kmer-classifier-output-taxid-seq
Option to enable a taxid string column in the output file. (Default=false).
--kmer-classifier-db-to-taxid-json
Path to JSON file that maps database IDs to external taxids, names, and ranks.
--kmer-classifier-no-read-output
Option to not create individual read output. (Default=false).
--kmer-classifier-no-taxid-counts
Option to not write taxid count output file. (Default=false).
--kmer-classifier-protein-input
Option to indicate protein query sequences. To use this option, the reference sequence database MUST be of protein sequences. (Default=false).
--kmer-classifier-ncpus
Option to set the number of CPUs available for processing.
1
Read index
integer
2
Read name
string
3
Taxid the read classified to
integer
4
Maximum number of contiguous kmers that classified to this taxid
integer
5
Score assigned to the classification
integer
6
Number of kmers that classified to this taxid
integer
7
Read duplication count
integer
8
Name associated with taxid, if given with --kmer-classifier-db-to-taxid-json
string
9
Taxonomic rank associated with taxid, if given with --kmer-classifier-db-to-taxid-json
string
10
Taxid that each kmer classified to (is output when the --kmer-classifier-output-taxid-seq flag is set)
list of integers separated by commas
11
Read sequence (is output when the the --kmer-classifier-output-read-seq flag is set)
string
db_taxid
Identifier for this taxid used internally in the database
integer
duplicity
Ratio of total number of kmers from reads assigned to this taxid compared to the number of distinct kmers from reads assigned to this taxid
float
distinct_coverage
Percent of kmers in the database assigned to this taxid that are covered by kmers in the reads assigned to this taxid
integer
read_count
Number of reads that classified to this taxid
integer
total_kmer_count
Number of kmers that classified to this taxid
integer
distinct_kmer_count
Number of distinct kmers that classified to this taxid
integer
cumulative_read_count
Cumulative number of reads assigned to this taxid and its taxonomic descendants
integer
taxid
Taxid
integer
name
Name associated with the taxid, if given with --kmer-classifier-db-to-taxid-json
string
rank
Taxonomic rank of the taxid, if given with --kmer-classifier-db-to-taxid-json
string
taxid_distinct_kmer_count
Number of distinct kmers assigned to this taxid from the reference sequences
string
probability_present
Not in use
float
Required Inputs |
| Enables the Kmer Classifier Database Builder. (Default=false). |
| Headerless, tab-delimited file where each line is (1) the path to a reference fasta file and (2) the associated taxid. When using --kmer-class-db-builder-taxids-as-seq-name, the second column is required but ignored. |
| Prefix for all output files. |
| Directory for all output files. |
| Kmer length (Range: [4, 31]). |
| Gmer length (must be >= kmer length. Range: [4, 64]). |
Optional Inputs |
| .tri file with nodes in the taxonomic tree for a classifier database (not required if building a binner database). Headerless, tab-delimited file where each line has (1) the child node taxid and (2) the parent node taxid. Root of tree must be 1 and have a parent of 0. |
| Set to indicate input sequences are protein sequences. (Default=false). |
| File with taxids to keep. If set, any kmers with taxids not in this file will be excluded from database. |
| Set to build a binner database with this number of categories. Max is 25 categories, assumes categories are from 2^0..2^n sequentially. The categories take the place of taxids in the input file. |
| Set to build classification database that saves all kmers / taxids / weights. |
| Cutoff that excludes k-mers that are found in more than cutoff number of taxids when building a database using --kmer-class-db-builder-save-weights. Helps speed up classification. (Default=1000). |
| Number of bits to mask in kmer before building / searching. (Default=7). |
| Option to set the number of CPUs available for processing. |
| Set to output number of kmers in each minimizer bucket. (Default=false). |
| Set to include kmers with lowercase bases (usually repeatmasked). (Default=false). |
| Set to indicate that the reference fastas listed in the input file have taxids as sequence name. In this case, the second column of the input file is ignored. (Default=false). |
The Explify Analysis Pipeline offers a dedicated informatics solution with flexible analysis options for the following Illumina Infectious Disease and Microbiology target-capture enrichment panel kits: the Illumina Respiratory Pathogen ID/AMR Enrichment Panel Kit (RPIP), Illumina Urinary Pathogen ID/AMR Enrichment Panel Kit (UPIP), and Illumina Viral Surveillance Panel V2 Kit (VSP V2). The application delivers easy-to-use, powerful secondary analysis of Illumina sequencing data, with workflows for sample QC, viral WGS (whole-genome sequencing), pathogen detection and quantification, and antimicrobial resistance (AMR) marker profiling. It also supports custom reference sequence analysis.
RPIP: Target-capture enrichment of >280 RNA and DNA respiratory pathogens, including SARS-CoV-2, Influenza viruses, Respiratory syncytial virus, Mycobacterium and Legionella species, and >4000 AMR markers.
UPIP: Target-capture enrichment of >170 genitourinary pathogens, including fastidious, slow-growing, and anaerobic uropathogens, sexually transmitted microorganisms, and >4000 bacterial AMR markers.
VSP V2: Target-capture enrichment for whole-genome sequencing (WGS) of 200 RNA and DNA viruses prioritized as high-risk to public health, zoonotic surveillance, and biotech, and >200 viral AMR markers.
Custom: Analyze FASTQ/FASTA read files with a custom reference sequence database.
Note that samples enriched using the Illumina Respiratory Virus Oligo Panel/Respiratory Virus Enrichment Kit (RVOP/RVEK) and Viral Surveillance Panel Kit (VSP) can also be analyzed using the Explify Analysis Pipeline and VSPv2 database.
| Option | Description |
|---|
Applies to: --explify-sample-list
The sample input list is a column-formatted file with tab separations between the columns (i.e., a .tsv file).
Notes:
The SampleID values must be unique.
BatchID and RunID are to help users track and manage sample analyses. Often the BatchID is used to track libraries that were prepared together, and the RunID is used to track sequencing runs. They can also be left blank.
The ControlFlag value can be POS, NEG, BLANK, or left empty.
POS is used to indicate a positive control sample.
NEG is used to indicate a negative control sample.
BLANK is used to indicate a blank control sample (e.g. buffer only).
If there are multiple FASTQ files, they are tab delimited.
Please be very careful when editing tsv files. Some editors replace tabs with spaces without alerting the user.
Applies to: --explify-internal-control, --explify-internal-control-concentration
The user may specify one of the internal controls listed below. If NONE is specified, the internal control concentration is ignored. These are case-sensitive and must be input exactly as they appear:
Allobacillus halotolerans
Armored RNA Quant Internal Process Control
Enterobacteria phage T7 (This is the default)
Escherichia virus MS2
Escherichia virus Qbeta
Escherichia virus T4
Imtechella halotolerans
Phocid alphaherpesvirus 1
Phocine morbillivirus
Truepera radiovictrix
NONE
The internal control concentration is an integer representing the number of copies/mL of sample for the internal control.
Applies to: --explify-ref-db-dir, --explify-test-panel-name, --explify-test-panel-version, --explify-load-db-ram,--explify-custom-ref-fasta, --explify-custom-ref-bed
An Explify Reference Database is required to run the Explify Analysis Pipeline in DRAGEN. The databases are stored remotely and must be downloaded prior to running an analysis. The database download script provided to facilitate the download is described below.
Prior to downloading the databases, create a directory that will be dedicated to storing them. It is recommended that the directory be on a disk with at least 150 GB of free space. The path to this directory will be used for the -d parameter when the download script is run in subsequent steps: "explify-databases/" is used in the examples below.
Download and management of Explify reference databases is handled by a shell script. The script can be downloaded with the following command:
The search subcommand can be used to list what databases can be downloaded:
The -d argument is the base directory used for storage of the databases
Optionally, when a test panel name is specified with the -p argument, the results will be limited to that panel
Optionally, setting the -n argument will filter the search to databases that have not already been downloaded
The download subcommand is used to download the database files for a test panel:
The -d argument is the base directory used for storage of the databases
The -p argument is the test panel name
The -v argument is the test panel version
The -n argument is the number of CPUs that can be used to download the files (defaults to 1)
Additional notes:
In this example, after the UPIP-8.4.0 files are downloaded, additional required files will be downloaded to a subdirectory named "common"
After the files are downloaded, their checksums will be automatically checked
Due to the size of some of the files, this command will take some time. It is best to run it via screen or nohup
The list subcommand is used to view the databases that have already been downloaded:
The -d argument is the base directory used for storage of the databases
Optionally, when a test panel name is specified with the -p argument, the results will be limited to that panel
The download subcommand will automatically check the file checksums after download. The check subcommand can also be used on its own to check the files:
The -d argument is the base directory used for storage of the databases
The -p argument is the test panel name
The -v argument is the test panel version
The -n argument is the number of CPUs that can be used to download the files (defaults to 1)
Assume the Explify database distributable, when unpacked, has a root directory name of /explify-databases. The database files will be organized in this root directory first by test panel type, then by test panel version:
To run an analysis with RPIP 6.3.0, for example, the following inputs would be needed:
The Explify Analysis Pipeline will use these inputs to navigate to the specified database location, namely /explify-databases/RPIP/6.3.0.
If the databases are stored on a normal file system, it is recommended that you set --explify-load-db-ram=true. This will tell the Explify Analysis Pipeline to load the databases into memory for faster analysis. It is also allowable to store the databases on a RAM disk, which reduces load time over many Explify Analysis Pipeline runs. In this case, it is recommended to set --explify-load-db-ram=false.
To use a Custom database, references are supplied through a FASTA file via --explify-custom-ref-fasta and an optional BED file via --explify-custom-ref-bed. Note that you must have downloaded the Custom database and set --explify-test-panel-name to "Custom", and set --explify-test-panel-version to the version you have downloaded. The supplied Custom Explify Reference Database is used by the Explify Analysis Pipeline to filter out host reads.
In the FASTA file, sequence names must be unique and should not contain any spaces. If there is any space in the FASTA header, the part before the first space is assumed to be the sequence name. It is recommended to use only the following in sequence names: alphabets, numbers, underscore (_), hyphen (-), parentheses ((,)), and period (.). Otherwise, the sequence names may appear different in the output.
The BED file must be tab-delimited with at least 4 columns:
chrom: the sequence name as it appears in the FASTA
chromStart: start position (always set to 0)
chromEnd: end position (sequence length)
genomeName: name of the genome, target, or microorganism the sequence belongs to (e.g. Monkeypox virus clade II)
segmentName (optional): the name of the segment or gene (e.g. Segment 4 (HA)). Set to 'Full' if the sequence is the full genome
Sequence names must match between the FASTA file and BED file, and the same set of sequences must appear in both files. If there are multiple viruses, their names should be unique. For example, if there are multiple Influenza genomes, they should not be labeled with the same virus name in the 4th column.
The BED file controls how sequences are labeled in the output JSON. If the custom reference FASTA file includes sequences from multiple segments, it is recommended to provide a BED file so that the segments are included under the results of that microorganism.
The output of the Explify Analysis Pipeline is a single ap.json file written to the specified output directory containing general metadata, version information, sample QC, microorganism, and AMR marker results, as well as detailed test information.
Top-Level Node
The top-level section of the output JSON contains general metadata and version information.
.qcReport Node
This section contains information about sample quality control (QC). The fields are relative to .qcReport
.qcReport.sampleComposition Node
This section contains information about the composition of the sample. The fields are relative to .qcReport.sampleComposition
.qcReport.internalControls Node
The value of the .qcReport.internalControls field is an array of objects containing name and RPKM information for each Internal Control. See the code block below for an example:
.userOptions Node
This section gives information about analysis options specified by the user. The fields are relative to .userOptions
.targetReport.microorganisms[] Node
The value of the .targetReport.microorganisms[] field is an array of objects containing information about detected microorganisms. The following table describes one .targetReport.microorganisms[] object. The fields are relative to .targetReport.microorganisms[]
.targetReport.microorganisms[].relatedMicroorganisms[] Node
The value of the .targetReport.microorganisms[].relatedMicroorganisms[] field is an array of objects containing information about genetically related microorganisms. The following table describes one .targetReport.microorganisms[].relatedMicroorganisms[] object. The fields are relative to .targetReport.microorganisms[].relatedMicroorganisms[]
.targetReport.microorganisms[].variants[] Node
The value of the .targetReport.microorganisms[].variants[] field is an array of objects containing information about viral variants for all VSPv2 viruses and select RPIP WGS viruses (SARS-CoV-2 & FluA/B/C). The following table describes one .targetReport.microorganisms[].variants[] object. The fields are relative to .targetReport.microorganisms[].variants[]
.targetReport.amrMarkers[] Node
The value of the .targetReport.amrMarkers[] field is an array of objects containing information about detected bacterial AMR markers. The following table describes one .targetReport.amrMarkers[] object. The fields are relative to .targetReport.amrMarkers[]
.targetReport.amrMarkers[].variants[] Node
The value of the .targetReport.amrMarkers[].variants[] field is an array of objects containing information about variants for bacterial AMR markers with "protein variant" or "rRNA variant" model types. The following table describes one .targetReport.amrMarkers[].variants[] object. The fields are relative to .targetReport.amrMarkers[].variants[]
.targetReport.customReferences[] Node
This section contains information about custom reference detection results and is only present for custom database analyses. When only a custom reference FASTA file is provided (no BED file), each .targetReport.customReferences[] object contains information for a single reference sequence. When both a FASTA and BED file are provided, each .targetReport.customReferences[] object contains information for a single genome/microorganism, which can be a collection of one or more reference sequences. The fields are relative to .targetReport.customReferences[]
.targetReport.customReferences[].consensusSequences[] Node
The value of the .targetReport.customReferences[].consensusSequences[] field is an array of objects containing majority consensus sequence information for a single custom reference sequence. When only a FASTA file is provided (no BED file), there will be only one object in the array. When both a FASTA and BED file are provided, there may be more than one object in the array. The fields are relative to .targetReport.customReferences[].consensusSequences[]
.targetReport.customReferences[].variants[] Node
The value of the .targetReport.customReferences[].variants[] field is an array of objects containing information about a single detected variant. The fields are relative to .targetReport.customReferences[].variants[]
| Field | Description |
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| Field | Description |
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| Field | Description |
|---|
Required Inputs |
| Enables the Explify Analysis Pipeline. (Default=false) |
| Prefix for all output files. |
| Directory for all output files. |
| Input sample list .tsv file with sample IDs, FASTQs, etc. |
| "RPIP", "UPIP", "VSPv2", "Custom". |
| Set to test panel version (e.g. "1.0.0"). |
| Path to root directory for Explify Database files. |
Optional Inputs |
| Area for temporary files. Size must be greater than size of all FASTQ files multiplied by 3. |
| Option to load database into RAM if not on ramdisk. (Default=false). |
| Option to turn off read QC on FASTQs before analysis. (Default=false). |
| Option to set internal control from an accepted list. (Default="Enterobacteria phage T7") |
| Option to set internal control concentration. (Default=12100000) |
| Option to set the number of CPUs available for processing. |
| Option to set sensitivity threshold. Range: 0 < Integer < 1000. Only valid for VSPv2. (Default=5). |
| Reference FASTA file. Required for Custom reference DBs. |
| Reference BED file. Optional for Custom reference DBs. |
.accession | Identifier used for the sample. |
.deploymentEnvironment | Environment in which the results were produced. |
.batchId | Identifier used for the batch of samples processed together. |
.analysisId | Identifier used for the analysis. |
.runId | Identifier used for the sequencing run. |
.controlFlag | Indicates whether the sample is a control. It is based on the ControlFlag field in the sample |
.dragenVersion | DRAGEN release version. |
.analysisPipelineVersion | Analysis Pipeline release version. |
.testType | Type of test panel ("RPIP", "UPIP", "VSPv2", "Custom"). |
.testVersion | Test panel release version. |
.testName | Name of the test panel, e.g. "Explify® Respiratory Pathogen ID/AMR Panel (RPIP) - Data Analysis Solution". |
.testUse | Test use. "For Research Use Only. Not for use in diagnostic procedures". |
.reportTime | Time the report was generated. |
.warnings | List of warnings encountered during the analysis. |
.errors | List of errors encountered during the analysis. |
.sampleQc | Sample QC information. |
.sampleQc.totalRawBases | Number of base pairs in sample before read QC processing. |
.sampleQc.totalRawReads | Number of reads in sample before read QC processing. |
.sampleQc.uniqueReads | Number of distinct reads in sample before read QC processing. |
.sampleQc.uniqueReadsProportion | Proportion of distinct reads in sample before read QC processing. |
.sampleQc.preQualityMeanReadLength | Average read length before read QC processing. |
.sampleQc.postQualityMeanReadLength | Average read length after read QC processing. |
.sampleQc.postQualityReads | Number of reads in sample after read QC processing. |
.sampleQc.postQualityReadsProportion | Proportion of post-quality reads in sample relative to total raw reads. |
.sampleQc.removedInDehostingReads | Number of host reads in sample removed during dehosting. |
.sampleQc.removedInDehostingReadsProportion | Proportion of host reads in sample removed relative to total raw reads. |
.sampleQc.entropy | Kmer entropy of reads after read QC processing. |
.sampleQc.gContent | Proportion of guanine (G) base calls in reads after read QC processing. |
.sampleQc.libraryQScore | Quality score of the library after read QC processing. |
.sampleQc.enrichmentFactor | Enrichment factor information (calculation requires detection of an appropriate Internal Control). |
.sampleQc.enrichmentFactor.value | Enrichment factor value reflecting how well targeted regions were enriched. |
.sampleQc.enrichmentFactor.category | Enrichment factor category: "poor", "fair", "good", or "not calculated". |
.readClassification | Proportion of reads classified to the following categories: |
.readClassification.targetedMicrobial | Targeted microbial. |
.readClassification.targetedInternalControl | Targeted Internal Control. |
.readClassification.untargeted | Untargeted. |
.readClassification.ambiguous | More than one category. |
.readClassification.unclassified | No category. |
.readClassification.lowComplexity | Low complexity. |
.targetedMicrobial | Proportion of targeted microbial reads classified to the following sub-categories: |
.targetedMicrobial.viral | Viral targeted. |
.targetedMicrobial.bacterial | Bacterial targeted. |
.targetedMicrobial.fungal | Fungal targeted. |
.targetedMicrobial.parasitic | Parasitic targeted. |
.targetedMicrobial.bacterialAmr | Bacterial AMR targeted. |
.untargeted | Proportion of untargeted reads classified to the following sub-categories: |
.untargeted.viral | Viral untargeted. |
.untargeted.bacterial | Bacterial untargeted. |
.untargeted.fungal | Fungal untargeted. |
.untargeted.parasitic | Parasitic untargeted. |
.untargeted.bacterialAmr | Bacterial AMR untargeted. |
.untargeted.internalControl | Internal Control untargeted. |
.untargeted.human | Human untargeted. |
.viral | Proportion of viral reads classified to the following categories: |
.viral.targeted | Viral targeted. |
.viral.untargeted | Viral untargeted. |
.viral.untargetedSubcategories | Proportion of viral untargeted reads classified to the following sub-categories: |
.viral.untargetedSubcategories.panel | Viral panel members. |
.viral.untargetedSubcategories.phage | Viral phage. |
.viral.untargetedSubcategories.other | Viral other (not a panel member or phage). |
.bacterial | Proportion of bacterial reads classified to the following categories: |
.bacterial.targeted | Bacterial targeted. |
.bacterial.untargeted | Bacterial untargeted. |
.bacterial.untargetedSubcategories | Proportion of bacterial untargeted reads classified to the following sub-categories: |
.bacterial.untargetedSubcategories.panel | Bacterial panel members. |
.bacterial.untargetedSubcategories.ribosomalDna | Bacterial ribosomal DNA (16S). |
.bacterial.untargetedSubcategories.plasmid | Bacterial plasmids. |
.bacterial.untargetedSubcategories.other | Bacterial other (not a panel member, ribosomal DNA, or plasmid). |
.fungal | Proportion of fungal reads classified to the following categories: |
.fungal.targeted | Fungal targeted. |
.fungal.untargeted | Fungal untargeted. |
.fungal.untargetedSubcategories | Proportion of fungal untargeted reads classified to the following sub-categories: |
.fungal.untargetedSubcategories.panel | Fungal panel members. |
.fungal.untargetedSubcategories.ribosomalDna | Fungal ribosomal DNA (18S). |
.fungal.untargetedSubcategories.other | Fungal other (not a panel member or ribosomal DNA). |
.parasitic | Proportion of parasitic reads classified to the following categories: |
.parasitic.targeted | Parasitic targeted. |
.parasitic.untargeted | Parasitic untargeted. |
.parasitic.untargetedSubcategories | Proportion of parasitic untargeted reads classified to the following sub-categories: |
.parasitic.untargetedSubcategories.panel | Parasitic panel members. |
.parasitic.untargetedSubcategories.ribosomalDna | Parasitic ribosomal DNA (18S). |
.parasitic.untargetedSubcategories.other | Parasitic other (not a panel member or ribosomal DNA). |
.human | Proportion of human reads classified to the following categories: |
.human.untargeted | Human untargeted. |
.human.untargetedSubcategories | Proportion of human untargeted reads classified to the following sub-categories: |
.human.untargetedSubcategories.ribosomalDna | Human ribosomal DNA. |
.human.untargetedSubcategories.codingSequence | Human coding sequence. |
.human.untargetedSubcategories.other | Human other (not ribosomal DNA or coding sequence). |
.internalControl | Proporition of Internal Control reads classified to the following categories: |
.internalControl.targeted | Internal Control targeted. |
.internalControl.untargeted | Internal Control untargeted. |
.microbialAndInternalControl | Proportion of Microbial and Internal Control reads classified to the following categories: |
.microbialAndInternalControl.targeted | Microbial and Internal Control targeted. |
.microbialAndInternalControl.untargeted | Microbial and Internal Control untargeted. |
.bacterialAmr | Proportion of bacterial AMR reads classified to the following categories: |
.bacterialAmr.targeted | Bacterial AMR targeted. |
.bacterialAmr.untargeted | Bacterial AMR untargeted. |
.quantitativeInternalControlName | The quantitative Internal Control used for microorganism absolute quantification (recommendation: Enterobacteria phage T7). |
.quantitativeInternalControlConcentration | The quantitative Internal Control concentration (recommendation: 1.21 x 10^7 copies/mL of sample). |
.readQcEnabled | Boolean field that indicates whether read QC (trimming and filtering based on quality and read length) was enabled. |
.readClassificationSensitivity | (VSPv2 only) Sensitivity threshold for classifying reads. Determines whether alignment should proceed for a microorganism and/or reference sequence. |
.class | Microorganism class ("viral", "bacterial", "fungal", "parasite") |
.name | Name of microorganism. |
.coverage | Proportion of targeted microorganism reference sequence bases that appear in sample sequencing reads. |
.ani | Average nucleotide identity of consensus sequence to targeted microorganism reference sequences. |
.medianDepth | Median depth of sample sequencing reads aligned to targeted microorganism reference sequences, indicating the median number of times each targeted microorganism reference sequence base appears in sample sequencing reads. |
.condensedDepthVector | Read depth across the targeted microorganism reference sequences, condensed to 256 bins. |
.rpkm | Normalized representation of the number of sample sequencing reads aligned to targeted microorganism reference sequences (targeted Reads mapped Per Kilobase of targeted sequence per Million quality-filtered reads). |
.alignedReadCount | Number of sample sequencing reads that aligned to targeted microorganism reference sequences. |
.kmerReadCount | (UPIP only) Number of sample sequencing reads classified to targeted microorganism reference sequences. |
.absoluteQuantityRatio | Numerical absolute quantification value. |
.absoluteQuantityRatioFormatted | Formatted absolute quantification value with units. |
.phenotypicGroup | Grouping indicating general association with normal flora, colonization, or contamination from the environment or other sources, as well as general association with disease. |
.associatedAmrMarkers | (Bacteria only) Information about the bacterial AMR markers associated with the microorganism. |
.associatedAmrMarkers.applicable | Boolean indicating whether one or more bacterial AMR markers are associated with the microorganism. |
.associatedAmrMarkers.detected | List of detected bacterial AMR markers associated with the microorganism. |
.associatedAmrMarkers.predicted | List of predicted bacterial AMR markers associated with the microorganism. |
.consensusGenomeSequences | (RPIP/VSPv2 viruses only) Information about the majority consensus genome (or segment) sequence. |
.consensusGenomeSequences.sequence | Consensus genome (or segment) sequence bases. |
.consensusGenomeSequences.referenceAccession | Accession of the reference genome (or segment) sequence. |
.consensusGenomeSequences.referenceDescription | Description of the reference genome (or segment) sequence. |
.consensusGenomeSequences.referenceLength | Length of the reference genome (or segment) sequence. |
.consensusGenomeSequences.maximumAlignmentLength | Longest contiguous alignment between consensus sequence and reference genome (or segment) sequence. |
.consensusGenomeSequences.maximumGapLength | Longest contiguous alignment gap (insertion or deletion) between consensus sequence and reference genome (or segment) sequence. |
.consensusGenomeSequences.maximumUnalignedLength | Longest section of the reference genome (or segment) sequence not aligned to by consensus sequence. |
.consensusGenomeSequences.coverage | Proportion of reference genome (or segment) sequence bases that appear in sample sequencing reads. |
.consensusGenomeSequences.ani | Average nucleotide identity of consensus sequence to reference genome (or segment) sequence. |
.consensusGenomeSequences.alignedReadCount | Number of sample sequencing reads that aligned to reference genome (or segment) sequence. |
.consensusGenomeSequences.medianDepth | Median depth of sample sequencing reads aligned to reference genome (or segment) sequence, indicating the median number of times each reference genome (or segment) sequence base appears in sample sequencing reads. |
.consensusGenomeSequences.targetAnnotation | List of targeted region annotations for the reference genome (or segment) sequence. Each annotation is a JSON object with the following fields: start (int), end (int), strand (string: "+", "-"), target_name (string), type (string). |
.consensusGenomeSequences.condensedDepthVector | Read depth across the reference genome (or segment) sequence, condensed to 256 bins. |
.consensusTargetSequences | (RPIP viruses only) Information about the majority targeted region consensus sequences. |
.consensusTargetSequences.sequence | Consensus targeted region sequence bases. |
.consensusTargetSequences.name | Name of the targeted region. |
.consensusTargetSequences.referenceAccession | Accession of the targeted region reference sequence. |
.consensusTargetSequences.depthVector | Read depth across the targeted region reference sequence, not condensed. |
.predictionInformation | Information about microorganism prediction results. |
.predictionInformation.predictedPresent | Boolean indicating whether the microorganism passed its proprietary reporting logic algorithm. |
.predictionInformation.notes | List of notes about the prediction result. |
.predictionInformation.subpanels | List of pre-defined subpanels that the microorganism belongs to. |
.predictionInformation.relatedMicroorganisms | Array of objects with information about genetically related microorganisms. See below for details. |
.name | Name of related microorganism. |
.onPanel | Boolean indicating whether the related microorganism is a panel member. |
.kmerReadCount | (UPIP only) Number of sample sequencing reads classified to related microorganism reference sequences. |
.coverage | Proportion of related microorganism reference sequence bases that appear in sample sequencing reads. |
.ani | Average nucleotide identity of consensus sequence to related microorganism reference sequences. |
.alignedReadCount | Number of sample sequencing reads that aligned to related microorganism reference sequences. |
.referenceAccession | Accession of reference genome (or segment) sequence used for variant calling. |
.segment | (Segmented viruses only) Segment number of reference segment sequence. |
.ntChange | Nucleotide change associated with variant. |
.referencePosition | Variant position in reference genome (or segment) sequence. |
.referenceAllele | Reference allele at variant position. |
.variantAllele | Variant allele. |
.depth | Variant depth, indicating the number of times variant allele appears in sample sequencing reads. |
.alleleFrequency | Frequency of variant allele in sample sequencing reads. |
.class | Microorganism class ("bacterial"). |
.cardModelType | Bacterial AMR marker model type in the Comprehensive Antibiotic Resistance Database (CARD) ("homolog", "protein variant", "rRNA variant"). |
.cardGeneFamily | Bacterial AMR marker family in the Comprehensive Antibiotic Resistance Database (CARD). |
.name | Bacterial AMR marker name. |
.cardName | Bacterial AMR marker name in the Comprehensive Antibiotic Resistance Database (CARD). |
.ncbiName | Bacterial AMR marker name in the National Center for Biotechnology Information (NCBI). |
.referenceAccession | Accession of the bacterial AMR marker reference sequence. |
.coverage | Proportion of bacterial AMR marker reference sequence residues that appear in sample sequencing reads (protein alignment for "homolog" and "protein variant" model types; nucleotide alignment for "rRNA variant" model type). |
.pid | Percent identity of consensus sequence aligned to bacterial AMR marker reference sequence (protein alignment for "homolog" and "protein variant" model types; nucleotide alignment for "rRNA variant" model type). |
.medianDepth | Median depth of sample sequencing reads aligned to bacterial AMR marker reference sequence, indicating the median number of times each bacterial AMR marker sequence residue appears in sample sequencing reads (protein alignment for "homolog" and "protein variant" model types; nucleotide alignment for "rRNA variant" model type). |
.rpkm | Normalized representation of the number of sample sequencing reads aligned to bacterial AMR reference sequence (protein alignment for "homolog" and "protein variant" model types; nucleotide alignment for "rRNA variant" model type). |
.alignedReadCount | Number of sample sequencing reads that aligned to bacterial AMR reference sequence (protein alignment for "homolog" and "protein variant" model types; nucleotide alignment for "rRNA variant" model type). |
.nucleotideConsensusSequence | Nucleotide consensus sequence bases. |
.proteinConsensusSequence | Protein consensus sequence bases. |
.nucleotideDepthVector | Read depth across the bacterial AMR marker nucleotide reference sequence, not condensed. |
.proteinDepthVector | Read depth across the bacterial AMR marker protein reference sequence, not condensed. |
.associatedMicroorganisms | Information about the microorganisms associated with the bacterial AMR marker. |
.associatedMicroorganisms.all | List of all microorganisms associated with the bacterial AMR marker. |
.associatedMicroorganisms.detected | List of detected microorganisms associated with the bacterial AMR marker. |
.associatedMicroorganisms.predicted | List of predicted microorganisms associated with the bacterial AMR marker. |
.predictionInformation | Information about bacterial AMR marker prediction results. |
.predictionInformation.predictedPresent | Boolean indicating whether the bacterial AMR marker passed its proprietary reporting logic algorithm. |
.predictionInformation.confidence | Confidence level of bacterial AMR marker prediction ("high", "medium", "low"). |
.predictionInformation.notes | List of notes about the prediction result. |
.category | Variant category ("Bacterial Variant; Known AMR"). |
.referenceSourceMicroorganism | Microorganism that reference sequence is associated with in NCBI. |
.comments | Comments about variant. |
.product | Protein product of gene. |
.ntChange | Nucleotide change associated with variant. |
.referencePosition | Variant position in reference sequence. |
.referenceAllele | Reference allele at variant position. |
.variantAllele | Variant allele. |
.depth | Variant depth, indicating the number of times variant allele appears in sample sequencing reads. |
.alleleFrequency | Frequency of variant allele in sample sequencing reads. |
.annotation | Type of change (e.g. "Nonsynonymous Variant"). |
.aaChange | Amino acid change associated with variant. |
.epistaticGroups | List of epistatic groups variant is associated with. |
.name | Name of custom reference sequence, accession or genome/microorgannism. |
.coverage | Proportion of custom reference sequence bases that appear in sample sequencing reads. |
.ani | Average nucleolotide identity of consensus sequence to custom reference sequence or, if specified, collection of one or more custom reference sequences. |
.medianDepth | Median depth of sample sequencing reads aligned to custom reference sequence or, if specified, collection of one or more custom reference sequences, indicating the med\ian number of times each custom reference sequence base appears in sample sequencing reads. |
.condensedDepthVector | Read depth across custom reference sequence or, if specified, collection of one or more custom reference sequences, condensed to 256 bins. |
.rpkm | Normalized number of sample sequencing reads aligned to custom reference sequence or, if specified, collection of one or more custom reference sequences (targeted Reads mapped Per Kilobase of targeted sequence per Million quality-filtered reads). |
.alignedReadCount | Number of sample sequencing reads that aligned to custom reference sequence or, if specified, collection of one or more custom reference sequences. |
.consensusSequences | Array of objects with information about each consensus sequence. |
.variants | Array of objects with information about variants detected in custom reference sequence or, if specified, collection of one or more custom reference sequences. |
.sequence | Majority consensus sequence bases. |
.referenceAccession | Accession of custom reference sequence. |
.referenceDescription | Description of custom reference sequence. |
.referenceLength | Length of custom reference sequence. |
.coverage | Proportion of custom reference sequence bases that appear in sample sequencing reads. |
.ani | Average nucleolotide identity of consensus sequence to custom reference sequence. |
.medianDepth | Median depth of sample sequencing reads aligned to custom reference sequence, indicating the median number of times each custom reference sequence base appears in sample sequencing reads. |
.depthVector | Read depth across custom reference sequence, not condensed. |
.alignedReadCount | Number of sample sequencing reads that aligned to custom reference sequence. |
.maximumAlignmentLength | Longest contiguous alignment between consensus sequence and custom reference sequence. |
.maximumGapLength | Longest contiguous alignment gap (insertion or deletion) between consensus sequence and custom reference sequence. |
.maximumUnalignedLength | Longest section of custom reference sequence not aligned to by consensus sequence. |
.ntChange | Nucleotide change associated with the variant. |
.referenceAccession | Accession of custom reference sequence used for variant calling. |
.referencePosition | Variant position in custom reference sequence. |
.referenceAllele | Reference allele at variant position. |
.variantAllele | Variant allele. |
.depth | Variant depth, indicating the number of times variant allele appears in sample sequencing reads. |
.alleleFrequency | Frequency of variant allele in sample sequencing reads. |