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ITSxpress: Software to rapidly trim the Internally transcribed spacer (ITS) region of FASTQ files

Project description

==================================================================================================
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Author
-------
* Adam R. Rivers, US Department of Agriculture, Agricultural Research Service


Introduction
-------------

The internally transcribed spacer region is a region between highly conserved the small
subunit (SSU) of rRNA and the large subunit (LSU) of the rRNA. In Eukaryotes it contains
the 5.8s genes and two variable length spacer regions. In amplicon sequening studies it is
common practice to trim off the conserved (SSU, 5,8S or LSU) regions. `Bengtsson-Palme
et al. (2013)`_ published software the software package ITSx_ to do this.

ITSxpress is designed to support the calling of exact sequence variants rather than OTUs_.
This newer method of sequence error-correction requires quality score data from each
sequence, so each input sequence must be trimmed. ITSXpress makes this possible by
taking FASTQ data, de-replicating the sequences then identifying the start and stop
sites using HMMSearch. Results are parsed and the trimmed files are returned. The ITS 1,
ITS2 or the entire ITS region including the 5.8s rRNA gene can be selected. ITSxpress
uses the hmm model from ITSx so results are comprable.


.. _`Bengtsson-Palme et al. (2013)`: https://doi.org/10.1111/2041-210X.12073
.. _ITSx: http://microbiology.se/software/itsx/
.. _OTUs: https://doi.org/10.1038/ismej.2017.119


Installation
-------------
ITSxpress can be installed from:

1. Preferred method - Bioconda (to be done):

.. code-block:: bash

conda install itsxpress

2. Pip (to be done):

.. code-block:: bash

pip install itsxpress


3. The Github repository: https://github.com/USDA-ARS-GBRU/itsxpress

.. code-block:: bash

git clone https://github.com/USDA-ARS-GBRU/itsxpress.git


Dependencies
-------------
The software requires Vsearch, BBtools, Hmmer and Biopython. Bioconda takes care of this
for you so it is the preferred installation method.


Usage
---------

-h, --help Show this help message and exit.

--fastq A ``.fastq``, ``.fq``, ``.fastq.gz`` or ``.fq.gz`` file. Interleaved
or not.

--single_end A flag to specify if the fastq file is inteleaved.
single-ended (not paired). Default is false.

--fastq2 A ``.fastq``, ``.fq``, ``.fastq.gz`` or ``.fq.gz`` file representing read 2, optional.

--outfile The trimmed Fastq file, if it ends in ``gz`` it will be gzipped.

--tempdir Specify the temp file directory.

--keeptemp Should intermediate files be kept?

--region Options : {ITS2, ITS1, ALL}

--taxa Select the taxonomic group sequenced: {Alveolata, Bryophyta,
Bacillariophyta, Amoebozoa, Euglenozoa, Fungi, Chlorophyta,
Rhodophyta, Phaeophyceae, Marchantiophyta, Metazoa, Microsporidia,
Oomycota, Haptophyceae, Raphidophyceae, Rhizaria, Synurophyceae,
Tracheophyta, Eustigmatophyceae, Apusozoa, Parabasalia}

--log Log file

--threads Number of processor threads to use


Examples
---------

Use case 1: Trimming the ITS2 region from a fungal amplicon sequencing dataset with
forward and reverse gzipped fastq files using two cpu threads.

.. code-block:: bash

itsxpress --fastq r1.fastq.gz --fastq2 r2.fastq.gz --region ITS2 --taxa Fungi \
--log logfile.txt --outfile trimmed_reads.fastq.gz --threads 2

ITSxpress can take gzipped or ungzipped fastq files and it can write gzipped or
ungzipped fastq files. It expects fastq files to end in : .fq, .fastq, .fq.gz or fastq.gz


Use case 2: Trimming the ITS2 region from a fungal amplicon sequencing dataset with
an interleaved gzipped fastq files using two cpu threads.

.. code-block:: bash

itsxpress --fastq interleaved.fastq.gz --region ITS2 --taxa Fungi \
--log logfile.txt --outfile trimmed_reads.fastq.gz --threads 2


Use case 3: Trimming the ITS2 region from a fungal amplicon sequencing dataset with
an interleaved gzipped fastq files using two cpu threads.

.. code-block:: bash

itsxpress --fastq single-end.fastq.gz --single_end --region ITS2 --taxa Fungi \
--log logfile.txt --outfile trimmed_reads.fastq.gz --threads 2

Single ended data is less common and may come from a dataset where the reads have already
been merged.

Use case 4: Trimming the ITS1 region from a Microsporidia amplicon sequencing dataset with
an interleaved gzipped fastq files using 40 cpu threads.

.. code-block:: bash

itsxpress --fastq interleaved.fastq.gz --region ITS1 --taxa Microsporidia \
--log logfile.txt --outfile trimmed_reads.fastq.gz --threads 40


License information
--------------------

This software is a work of the United States Department of Agriculture, Agricultural
Research Service. 17 U.S.C. Section 105 states that "Copyright protection under this
title is not available for any work of the United States Government". While I anticipate
that this work will be released under a CC0 public domain attribution, only the USDA ARS
Office of Technology transfer has the authority to make that determination.
Home-page: http://github.com/usda-ars-gbru/itsxpress
Author: Adam R. Rivers
Author-email: adam.rivers@ars.usda.gov
License: License :: CC0 1.0 Universal (CC0 1.0) Public Domain Dedication
Description: Rapidly trim sequences down to their Internally Transcribed Spacer (ITS) regions
Keywords: Amplicon sequencing fungal ITS
Platform: UNKNOWN
Classifier: Topic :: Scientific/Engineering :: Bio-Informatics
Classifier: Programming Language :: Python :: 3.6
Classifier: Programming Language :: Python :: 3.5
Classifier: Development Status :: 3 - Alpha
Requires-Python: >3.5

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