Metadata-Version: 2.1
Name: referenceseeker
Version: 1.7.1
Summary: ReferenceSeeker: rapid determination of appropriate reference genomes.
Home-page: https://github.com/oschwengers/referenceseeker
Author: Oliver Schwengers
Author-email: oliver.schwengers@computational.bio.uni-giessen.de
License: GPLv3
Project-URL: Bug Reports, https://github.com/oschwengers/referenceseeker/issues
Project-URL: Source, https://github.com/oschwengers/referenceseeker
Description: [![DOI](https://joss.theoj.org/papers/10.21105/joss.01994/status.svg)](https://doi.org/10.21105/joss.01994)
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        # ReferenceSeeker: rapid determination of appropriate reference genomes
        
        ## Contents
        
        - [Description](#description)
        - [Input & Output](#input-output)
        - [Installation](#installation)
          - [BioConda](#bioconda)
          - [GitHub](#github)
        - [Usage](#usage)
        - [Examples](#examples)
        - [Databases](#databases)
          - [RefSeq](#refseq-based)
          - [Custom](#custom-database)
        - [Dependencies](#dependencies)
        - [Citation](#citation)
        
        ## Description
        
        ReferenceSeeker determines closely related reference genomes following a scalable hierarchical approach combining an 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 potential reference genome candidates via Mash (Ondov et al. 2016). For resulting candidates ReferenceSeeker subsequently computes (bidirectional) 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 to take into account both genome coverage and identity.
        
        Custom databases can be built with local genomes. For further convenience, we provide pre-built databases with sequences from RefSeq (<https://www.ncbi.nlm.nih.gov/refseq>), GTDB and PLSDB copmrising the following taxa:
        
        - bacteria
        - archaea
        - fungi
        - protozoa
        - viruses
        
        as well as *plasmids*.
        
        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, however 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 conservation (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.
        
        ![Mash D vs. ANI / conDNA](mash-ani-cdna.mini.png?raw=true)
        
        ## Input & Output
        
        ### Input
        
        Path to a taxon database and a draft or finished genome in (zipped) fasta format:
        
        ```bash
        $ 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
        
        ```bash
        #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
        
        ```bash
        #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
        
        ReferenceSeeker can be installed via Conda and Git(Hub). In either case, a taxon database must be downloaded which we provide for download at Zenodo: [![DOI](https://zenodo.org/badge/DOI/10.5281/zenodo.3562004.svg)](https://doi.org/10.5281/zenodo.3562004)
        For more information have a look at [Databases](#databases).
        
        ### BioConda
        
        The preferred way to install and run ReferenceSeeker is [Conda](https://conda.io/docs/install/quick.html) using the [Bioconda](https://bioconda.github.io/) channel:
        
        ```bash
        $ conda install -c bioconda referenceseeker
        $ referenceseeker --help
        ```
        
        ### GitHub
        
        Alternatively, you can use this raw GitHub repository:
        
        1. install necessary Python dependencies (if necessary)
        2. clone the latest version of the repository
        3. install necessary 3rd party executables (Mash, MUMmer4)
        
        ```bash
        $ pip3 install --user biopython xopen
        $ git clone https://github.com/oschwengers/referenceseeker.git
        $ # install Mash & MUMmer
        $ ./referenceseeker/bin/referenceseeker --help
        ```
        
        ### Test
        
        To test your installation we prepared a tiny mock database comprising 4 `Salmonella spp` genomes and a query assembly (SRA: SRR498276) in the `tests` directory:
        
        ```bash
        $ git clone https://github.com/oschwengers/referenceseeker.git
        
          # GitHub installation
        $ ./referenceseeker/bin/referenceseeker referenceseeker/test/db referenceseeker/test/data/Salmonella_enterica_CFSAN000189.fasta
        
          # BioConda installation
        $ referenceseeker referenceseeker/test/db referenceseeker/test/data/Salmonella_enterica_CFSAN000189.fasta
        ```
        
        Expected output:
        
        ```bash
        #ID    Mash Distance    ANI    Con. DNA    Taxonomy ID    Assembly Status    Organism
        GCF_000439415.1    0.00003    100.00    99.55    1173427    complete    Salmonella enterica subsp. enterica serovar Bareilly str. CFSAN000189
        GCF_900205275.1    0.01522    98.61     83.13    90370      complete    Salmonella enterica subsp. enterica serovar Typhi
        ```
        
        ## Usage
        
        Usage:
        
        ```bash
        usage: referenceseeker [--crg CRG] [--ani ANI] [--conserved-dna CONSERVED_DNA]
                               [--unfiltered] [--bidirectional] [--help] [--version]
                               [--verbose] [--threads THREADS]
                               <database> <genome>
        
        Rapid determination of appropriate reference genomes.
        
        positional arguments:
          <database>            ReferenceSeeker database path
          <genome>              target draft genome in fasta format
        
        Filter options / thresholds:
          These options control the filtering and alignment workflow.
        
          --crg CRG, -r CRG     Max number of candidate reference genomes to pass kmer
                                prefilter (default = 100)
          --ani ANI, -a ANI     ANI threshold (default = 0.95)
          --conserved-dna CONSERVED_DNA, -c CONSERVED_DNA
                                Conserved DNA threshold (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/conserved DNA values
                                (default = False)
        
        Runtime & auxiliary options:
          --help, -h            Show this help message and exit
          --version, -V         show program's version number and exit
          --verbose, -v         Print verbose information
          --threads THREADS, -t THREADS
                                Number of used threads (default = number of available
                                CPU cores)
        ```
        
        ## Examples
        
        Installation:
        
        ```bash
        $ conda install -c bioconda referenceseeker
        $ wget https://zenodo.org/record/4415843/files/bacteria-refseq.tar.gz
        $ tar -xzf bacteria-refseq.tar.gz
        $ rm bacteria-refseq.tar.gz
        ```
        
        Simple:
        
        ```bash
        $ # referenceseeker <REFERENCE_SEEKER_DB> <GENOME>
        $ referenceseeker bacteria-refseq/ genome.fasta
        ```
        
        Expert: verbose output and increased output of candidate reference genomes using a defined number of threads:
        
        ```bash
        $ # referenceseeker --crg 500 --verbose --threads 8 <REFERENCE_SEEKER_DB> <GENOME>
        $ referenceseeker --crg 500 --verbose --threads 8 bacteria-refseq/ genome.fasta
        ```
        
        ## Databases
        
        ReferenceSeeker depends on databases comprising taxonomic genome informations as well as kmer hash profiles for each entry.
        
        ### Pre-built
        
        We provide pre-built databases based on public genome data hosted at Zenodo: [![DOI](https://zenodo.org/badge/DOI/10.5281/zenodo.4415843.svg)](https://doi.org/10.5281/zenodo.4415843) :
        
        #### RefSeq
        
        release: 205 (2021-04-01)
        
        | Taxon | URL | # Genomes | Size |
        | :---: | --- | ---: | :---: |
        | bacteria | <https://zenodo.org/record/4415843/files/bacteria-refseq.tar.gz> | 30,941 | 40 Gb |
        | archaea | <https://zenodo.org/record/4415843/files/archaea-refseq.tar.gz> | 606 | 553 Mb |
        | fungi | <https://zenodo.org/record/4415843/files/fungi-refseq.tar.gz> | 347 | 3.3 Gb |
        | protozoa | <https://zenodo.org/record/4415843/files/protozoa-refseq.tar.gz> | 88 | 1.1 Gb |
        | viruses | <https://zenodo.org/record/4415843/files/viral-refseq.tar.gz> | 10,339 | 730 Mb |
        
        #### GTDB
        
        release: v95 (2021-01-06)
        
        | Taxon | URL | # Genomes | Size |
        | :---: | --- | ---: | :---: |
        | bacteria | <https://zenodo.org/record/4415843/files/bacteria-gtdb.tar.gz> | 30,238 | 34 Gb |
        | archaea | <https://zenodo.org/record/4415843/files/archaea-gtdb.tar.gz> | 1,672 | 1.1 Gb |
        
        #### Plasmids
        
        In addition to the genome based databases, we provide the following plasmid databases based on RefSeq and PLSDB:
        
        | DB | URL | # Plasmids | Size |
        | :---: | --- | ---: | :---: |
        | RefSeq | <https://zenodo.org/record/4415843/files/plasmids-refseq.tar.gz> | 32,611 | 1.1 Gb |
        | PLSDB | <https://zenodo.org/record/4415843/files/plasmids-plsdb.tar.gz> | 27,393 | 1.1 Gb |
        
        ### Custom database
        
        If above mentiond RefSeq based databases do not contain sufficiently-close related genomes or are just too large, ReferenceSeeker provides auxiliary commands in order to either create databases from scratch or to expand existing ones. Therefore, a second executable `referenceseeker_db` accepts `init` and `import` subcommands:
        
        Usage:
        
        ```bash
        usage: referenceseeker_db [--help] [--version] {init,import} ...
        
        Rapid determination of appropriate reference genomes.
        
        positional arguments:
          {init,import}  sub-command help
            init         Initialize a new database
            import       Add a new genome to database
        
        Runtime & auxiliary options:
          --help, -h     Show this help message and exit
          --version, -V  show program's version number and exit
        ```
        
        If a new database should be created, use `referenceseeker_db init`:
        
        ```bash
        usage: referenceseeker_db init [-h] [--output OUTPUT] --db DB
        
        optional arguments:
          -h, --help            show this help message and exit
          --output OUTPUT, -o OUTPUT
                                output directory (default = current working directory)
          --db DB, -d DB        Name of the new ReferenceSeeker database
        ```
        
        This new database or an existing one can be used to import genomes in Fasta, GenBank or EMBL format:
        
        ```bash
        usage: referenceseeker_db import [-h] --db DB --genome GENOME [--id ID]
                                         [--taxonomy TAXONOMY]
                                         [--status {complete,chromosome,scaffold,contig}]
                                         [--organism ORGANISM]
        
        optional arguments:
          -h, --help            show this help message and exit
          --db DB, -d DB        ReferenceSeeker database path
          --genome GENOME, -g GENOME
                                Genome path [Fasta, GenBank, EMBL]
          --id ID, -i ID        Unique genome identifier (default sequence id of first
                                record)
          --taxonomy TAXONOMY, -t TAXONOMY
                                Taxonomy ID (default = 12908 [unclassified sequences])
          --status {complete,chromosome,scaffold,contig}, -s {complete,chromosome,scaffold,contig}
                                Assembly level (default = contig)
          --organism ORGANISM, -o ORGANISM
                                Organism name (default = "NA")
        ```
        
        ## Dependencies
        
        ReferenceSeeker needs the following dependencies:
        
        - Python (3.8, 3.9), Biopython (>=1.78), xopen(>=1.1.0)
        - Mash (2.3) <https://github.com/marbl/Mash>
        - MUMmer (4.0.0-beta2) <https://github.com/gmarcais/mummer>
        
        ReferenceSeeker has been tested against aforementioned versions.
        
        ## Citation
        
        > Schwengers et al., (2020). ReferenceSeeker: rapid determination of appropriate reference genomes. Journal of Open Source Software, 5(46), 1994, https://doi.org/10.21105/joss.01994
        
Keywords: bioinformatics,ngs,wgs,microbial genomics
Platform: UNKNOWN
Classifier: Operating System :: POSIX :: Linux
Classifier: Programming Language :: Python :: 3 :: Only
Classifier: Development Status :: 5 - Production/Stable
Classifier: License :: OSI Approved :: GNU General Public License v3 (GPLv3)
Classifier: Topic :: Scientific/Engineering :: Bio-Informatics
Classifier: Environment :: Console
Classifier: Intended Audience :: Science/Research
Classifier: Natural Language :: English
Requires-Python: >=3.8
Description-Content-Type: text/markdown
