q2-thapbi-pict 0.0.1

About

Qiime2 is a microbiome bioinformatics platform with integrated data provenance tracking and a plugin system allowing modular construction of analysis and visualisation pipelines.

THAPBI PICT is a sequence based diagnostic/profiling tool started under the UK funded Tree Health and Plant Biosecurity Initiative (THAPBI) Phyto-Threats project. PICT stood for Phytophthora ITS1 Classification Tool, reflecting the initial application focus.

With appropriate primer settings and a custom database of full length markers, THAPBI PICT can be applied to other organisms and/or barcode marker sequences. It requires overlapping paired-end Illumina reads which can be merged to cover the full amplicon marker. Longer markers or fragmented amplicons are not supported. Internally it works by tracking unique amplicon sequence variants (ASVs), using MD5 checksums as identifiers.

The THAPBI PICT tool provides a command line interface including a pipeline from demultiplexed FASTQ files and optional metadata as TSV files though to ASV tally tables with taxonomic classification as TSV, Excel or BIOM files.

This repository is for q2-thapbi-pict, a plugin to call some of the THAPBI PICT functionality from within Qiime2.

Installation

Qiime2 is not available natively on Windows, instead they recommend using the Windows Subsystem for Linux (WSL). Follow the Qiime2 installation instructions using the Conda packaging system.

1. Activate your Qiime2 conda environment.

2. Install the paired read merging tool Flash used by THAPBI PICT using conda:

   $ conda install -c conda-forge flash

3. Install the THAPBI PICT plugin for Qiime2 (and its Python dependencies which include THAPBI PICT and CutAdapt) using pip:

   $ pip install q2-thapbi-pict

   This will fail ERROR: No matching distribution found for q2-types... or ERROR: No matching distribution found for q2cli... if the Qiime2 conda environment is not activated.

Quick Start

Once installed, you should be able to run the tool at the command line using the Qiime2 command line interface (q2cli):

$ qiime info
System versions
Python version: 3.8.18
QIIME 2 release: 2024.2
QIIME 2 version: 2024.2.0
q2cli version: 2024.2.0

Installed plugins
...

This should list the THAPBI PICT plugin under “Installed plugins”. You should be able to run it and see basic instructions too:

$ qiime q2-thapbi-pict --help
Usage: qiime q2-thapbi-pict [OPTIONS] COMMAND [ARGS]...

  Description: This QIIME 2 plugin provides support for running some of the
  THAPBI PICT functionality.

...

This lists the available commands, each of which have their own help page:

$ qiime q2-thapbi-pict prepare-reads-sample-tally --help
Usage: qiime q2-thapbi-pict prepare-reads-sample-tally [OPTIONS]

  Takes paired (raw) FASTQ files demultiplexed per sample. Runs THAPBI PICT
  ...

You can confirm the plugin version like this:

$ qiime q2-thapbi-pict --version
QIIME 2 Plugin 'q2-thapbi-pict' version 0.0.1 (from package 'q2-thapbi-pict' version 0.0.1)

Documentation

The THAPBI PICT documentation is hosted by Read The Docs.

Citation

If you use THAPBI PICT in your work, please cite our PeerJ paper, and give details of the version and any non-default settings used in your methods:

Cock et al. (2023) “THAPBI PICT - a fast, cautious, and accurate metabarcoding analysis pipeline” PeerJ 11:e15648 https://doi.org/10.7717/peerj.15648

You can also cite the software specifically via Zenodo which offers version specific DOIs as well as https://doi.org/10.5281/zenodo.4529395 which is for the latest version.

Qiime2 helps by tracking the citations for all the tools your analysis uses. This should include any tools used via plugins, such as Flash and CutAdapt.