ReferenceSeeker: rapid determination of appropriate reference genomes.
Project description
ReferenceSeeker: rapid determination of appropriate reference genomes.
Contents
Description
ReferenceSeeker determines closely related reference genomes from RefSeq (https://www.ncbi.nlm.nih.gov/refseq) following a scalable hierarchical approach combining an ultra-fast kmer profile-based database lookup of candidate reference genomes and subsequent computation of specific average nucleotide identity (ANI) values for the rapid determination of suitable reference genomes.
ReferenceSeeker computes kmer-based genome distances between a query genome and and a database built on RefSeq genomes via Mash (Ondov et al. 2016). Therefore, only complete genomes or those stated as 'representative' or 'reference' genome are included. ReferenceSeeker offers pre-built databases for a broad spectrum of microbial taxonomic groups, i.e. bacteria, archaea, fungi, protozoa and viruses. For resulting candidates ReferenceSeeker subsequently computes ANI values picking genomes meeting community standard thresholds by default (ANI >= 95 % & conserved DNA >= 69 %) (Goris, Konstantinos et al. 2007) ranked by the product of ANI and conserved DNA values.
The reasoning for subsequent calculations of both ANI and conserved DNA values is that Mash distance values correlate well with ANI values for closely related genomes but the same is not true for conserved DNA values. A kmer fingerprint-based comparison alone cannot distinguish if a kmer is missing due to a SNP, for instance or a lack of the kmer-comprising subsequence. As DNA conservancy (next to DNA identity) is very important for many kinds of analyses, e.g. reference based SNP detections, ranking potential reference genomes based on a mash distance alone is often not sufficient in order to select the most appropriate reference genomes. If desired, ANI and conserved DNA values can be computed bidirectionally.
Input & Output
Input:
Path to a taxon database and a draft or finished genome in fasta format:
$ referenceseeker ~/bacteria GCF_000013425.1.fna
Output:
Tab separated lines to STDOUT comprising the following columns:
Unidirectionally (query -> references):
- RefSeq Assembly ID
- Mash Distance
- ANI
- Conserved DNA
- NCBI Taxonomy ID
- Assembly Status
- Organism
#ID Mash Distance ANI Con. DNA Taxonomy ID Assembly Status Organism
GCF_000013425.1 0.00000 100.00 100.00 93061 complete Staphylococcus aureus subsp. aureus NCTC 8325
GCF_001900185.1 0.00002 100.00 99.89 46170 complete Staphylococcus aureus subsp. aureus HG001
GCF_900475245.1 0.00004 100.00 99.57 93061 complete Staphylococcus aureus subsp. aureus NCTC 8325 NCTC8325
GCF_001018725.2 0.00016 100.00 99.28 1280 complete Staphylococcus aureus FDAARGOS_10
GCF_003595465.1 0.00185 99.86 96.81 1280 complete Staphylococcus aureus USA300-SUR6
GCF_003595385.1 0.00180 99.87 96.80 1280 complete Staphylococcus aureus USA300-SUR2
GCF_003595365.1 0.00180 99.87 96.80 1280 complete Staphylococcus aureus USA300-SUR1
GCF_001956815.1 0.00180 99.87 96.80 46170 complete Staphylococcus aureus subsp. aureus USA300_SUR1
...
Bidirectionally (query -> references [QR] & references -> query [RQ]):
- RefSeq Assembly ID
- Mash Distance
- QR ANI
- QR Conserved DNA
- RQ ANI
- RQ Conserved DNA
- NCBI Taxonomy ID
- Assembly Status
- Organism
#ID Mash Distance QR ANI QR Con. DNA RQ ANI RQ Con. DNA Taxonomy ID Assembly Status Organism
GCF_000013425.1 0.00000 100.00 100.00 100.00 100.00 93061 complete Staphylococcus aureus subsp. aureus NCTC 8325
GCF_001900185.1 0.00002 100.00 99.89 100.00 99.89 46170 complete Staphylococcus aureus subsp. aureus HG001
GCF_900475245.1 0.00004 100.00 99.57 99.99 99.67 93061 complete Staphylococcus aureus subsp. aureus NCTC 8325 NCTC8325
GCF_001018725.2 0.00016 100.00 99.28 99.95 98.88 1280 complete Staphylococcus aureus FDAARGOS_10
GCF_001018915.2 0.00056 99.99 96.35 99.98 99.55 1280 complete Staphylococcus aureus NRS133
GCF_001019415.2 0.00081 99.99 94.47 99.98 99.36 1280 complete Staphylococcus aureus NRS146
GCF_001018735.2 0.00096 100.00 94.76 99.98 98.58 1280 complete Staphylococcus aureus NRS137
GCF_003354885.1 0.00103 99.93 96.63 99.93 96.66 1280 complete Staphylococcus aureus 164
...
Installation
Platon can be installed via Conda and Git(Hub).
In either case, a taxon database must be downloaded which we provide for download at Zenodo: For more information scroll to Databases.
Conda / BioConda
The preferred way to install and run ReferenceSeeker is Conda using the Bioconda channel:
$ conda install -c conda-forge -c bioconda -c defaults referenceseeker
$ referenceseeker --help
GitHub
Alternatively, you can use this raw GitHub repository:
- install necessary Python dependencies (if necessary)
- clone the latest version of the repository
- download and extract a databases
Example:
$ pip3 install --user biopython
$ git clone https://github.com/oschwengers/referenceseeker.git
$ ./referenceseeker/bin/referenceseeker --help
Usage
Usage:
usage: referenceseeker [-h] [--crg CRG] [--ani ANI]
[--conserved-dna CONSERVED_DNA] [--unfiltered]
[--bidirectional] [--verbose] [--threads THREADS]
[--version]
<database> <genome>
Rapid determination of appropriate reference genomes.
positional arguments:
<database> ReferenceSeeker database path
<genome> target draft genome in fasta format
optional arguments:
-h, --help show this help message and exit
--crg CRG, -r CRG max number of candidate reference genomes to pass kmer
prefilter (default = 100)
--ani ANI, -a ANI ANI threshold value (default = 0.95)
--conserved-dna CONSERVED_DNA, -c CONSERVED_DNA
Conserved DNA threshold value (default = 0.69)
--unfiltered, -u set kmer prefilter to extremely conservative values
and skip species level ANI cutoffs (ANI >= 0.95 and
conserved DNA >= 0.69
--bidirectional, -b Compute bidirectional ANI values (default = False)
--verbose, -v print verbose information
--threads THREADS, -t THREADS
number of threads to use (default = number of
available CPUs)
--version, -V show program's version number and exit
Examples
Simple:
$ # referenceseeker <REFERENCE_SEEKER_DB> <GENOME>
$ referenceseeker bacteria/ genome.fasta
Expert: verbose output and increased output of candidate reference genomes using a defined number of threads:
$ # referenceseeker --crg 500 --verbose --threads 8 <REFERENCE_SEEKER_DB> <GENOME>
$ referenceseeker --crg 500 --verbose --threads 8 bacteria/ genome.fasta
Databases
ReferenceSeeker depends on custom databases based on reference, representative as well as complete NCBI RefSeq genomes comprising kmer hash profiles and taxonomic information. We provide the following pre-built databases based on RefSeq 2019-07-02 via :
Taxon | URL | # Genomes | Size Zipped | Size Unzipped |
---|---|---|---|---|
bacteria | https://zenodo.org/record/3562005/files/bacteria.tar.gz?download=1 | 18,229 | 22 Gb | 71 Gb |
archaea | https://zenodo.org/record/3562005/files/archaea.tar.gz?download=1 | 417 | 364 Mb | 1.2 Gb |
fungi | https://zenodo.org/record/3562005/files/fungi.tar.gz?download=1 | 288 | 2.6 Gb | 8 Gb |
protozoa | https://zenodo.org/record/3562005/files/protozoa.tar.gz?download=1 | 88 | 1 Gb | 3.4 Gb |
viral | https://zenodo.org/record/3562005/files/viral.tar.gz?download=1 | 9,264 | 608 Mb | 835 Mb |
Updated database versions reflecting the latest RefSeq versions can be built with a shell script and nextflow pipeline.
Download and install Nextflow:
$ curl -fsSL get.nextflow.io | bash
Build database:
$ export REFERENCE_SEEKER_HOME=<REFERENCE_SEEKER_DIR>
$ sh $REFERENCE_SEEKER_HOME/build-db.sh <DB_TYPE_OPTION>
List of available database options:
$ sh build-db.sh
-b (bacteria)
-a (archaea)
-v (viral)
-f (fungi)
-p (protozoa)
Dependencies
ReferenceSeeker needs the following dependencies:
- Python (3.5.2), Biopython (1.71)
- Mash (2.2) https://github.com/marbl/Mash
- MUMmer (4.0.0-beta2) https://github.com/gmarcais/mummer
ReferenceSeeker has been tested against aforementioned versions.
Citation
ReferenceSeeker: rapid determination of appropriate reference genomes. Oliver Schwengers, Torsten Hain, Trinad Chakraborty, Alexander Goesmann. bioRxiv 863621; doi: https://doi.org/10.1101/863621
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