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#!/usr/bin/env python3
from __future__ import with_statement
__author__ = ('Nicola Segata (nicola.segata@unitn.it), '
'Duy Tin Truong, '
'Francesco Asnicar (f.asnicar@unitn.it), '
'Francesco Beghini (francesco.beghini@unitn.it)')
__version__ = '2.9.22'
__date__ = '14 Oct 2019'
import sys
import os
import stat
import re
import time
import tarfile
try:
import numpy as np
except ImportError:
sys.stderr.write("Error! numpy python library not detected!!\n")
sys.exit(1)
p2 = float(sys.version_info[0]) < 3.0
if p2:
DEVNULL = open(os.devnull, 'wb')
import cPickle as pickle
from urllib import urlretrieve
else:
from subprocess import DEVNULL
import pickle
from urllib.request import urlretrieve
import tempfile as tf
import argparse as ap
import subprocess as subp
from collections import defaultdict as defdict
import bz2
import itertools
from distutils.version import LooseVersion
import pickle
from glob import glob
import hashlib
# set the location of the database download url
DATABASE_DOWNLOAD = "https://bitbucket.org/biobakery/metaphlan2/downloads/"
# get the directory that contains this script
metaphlan2_script_install_folder = os.path.dirname(os.path.abspath(__file__))
# get the default database folder
DEFAULT_DB_FOLDER = os.path.join(metaphlan2_script_install_folder, "metaphlan_databases")
#**********************************************************************************************
# Modification of Code : *
# Modified the code so instead of using the current clade IDs, which are numbers, we will *
# use the clade_names *
# Users reported the biom output is invalid and also the IDs were changing from run to *
# run. *
# George Weingart 05/22/2017 george.weingart@mail.com *
#**********************************************************************************************
#*************************************************************
#* Imports related to biom file generation *
#*************************************************************
try:
import biom
import biom.table
except ImportError:
sys.stderr.write("Warning! Biom python library not detected!"
"\n Exporting to biom format will not work!\n")
try:
import json
except ImportError:
sys.stderr.write("Warning! json python library not detected!"
"\n Exporting to biom format will not work!\n")
tax_units = "kpcofgst"
def remove_prefix(text):
return re.sub(r'^[a-z]__', '', text)
if float(sys.version_info[0]) < 3.0:
def read_and_split(ofn):
return (l.strip().split('\t') for l in ofn)
def read_and_split_line(line):
return line.strip().split('\t')
else:
def read_and_split(ofn):
return (l.decode('utf-8').strip().split('\t') for l in ofn)
def read_and_split_line(line):
return line.decode('utf-8').strip().split('\t')
def plain_read_and_split(ofn):
return (l.strip().split('\t') for l in ofn)
def plain_read_and_split_line(l):
return l.strip().split('\t')
if float(sys.version_info[0]) < 3.0:
def mybytes(val):
return val
else:
def mybytes(val):
return bytes(val, encoding='utf-8')
def read_params(args):
p = ap.ArgumentParser( description=
"DESCRIPTION\n"
" MetaPhlAn version "+__version__+" ("+__date__+"): \n"
" METAgenomic PHyLogenetic ANalysis for metagenomic taxonomic profiling.\n\n"
"AUTHORS: "+__author__+"\n\n"
"COMMON COMMANDS\n\n"
" We assume here that metaphlan2.py is in the system path and that mpa_dir bash variable contains the\n"
" main MetaPhlAn folder. Also BowTie2 should be in the system path with execution and read\n"
" permissions, and Perl should be installed)\n\n"
"\n========== MetaPhlAn 2 clade-abundance estimation ================= \n\n"
"The basic usage of MetaPhlAn 2 consists in the identification of the clades (from phyla to species and \n"
"strains in particular cases) present in the metagenome obtained from a microbiome sample and their \n"
"relative abundance. This correspond to the default analysis type (-t rel_ab).\n\n"
"* Profiling a metagenome from raw reads:\n"
"$ metaphlan2.py metagenome.fastq --input_type fastq -o profiled_metagenome.txt\n\n"
"* You can take advantage of multiple CPUs and save the intermediate BowTie2 output for re-running\n"
" MetaPhlAn extremely quickly:\n"
"$ metaphlan2.py metagenome.fastq --bowtie2out metagenome.bowtie2.bz2 --nproc 5 --input_type fastq -o profiled_metagenome.txt\n\n"
"* If you already mapped your metagenome against the marker DB (using a previous MetaPhlAn run), you\n"
" can obtain the results in few seconds by using the previously saved --bowtie2out file and \n"
" specifying the input (--input_type bowtie2out):\n"
"$ metaphlan2.py metagenome.bowtie2.bz2 --nproc 5 --input_type bowtie2out -o profiled_metagenome.txt\n\n"
"* bowtie2out files generated with MetaPhlAn2 versions below 2.9 are not compatibile.\n"
" Starting from MetaPhlAn2 2.9, the BowTie2 ouput now includes the size of the profiled metagenome.\n"
" If you want to re-run MetaPhlAn2 using these file you should provide the metagenome size via --nreads:\n"
"$ metaphlan2.py metagenome.bowtie2.bz2 --nproc 5 --input_type bowtie2out --nreads 520000 -o profiled_metagenome.txt\n\n"
"* You can also provide an externally BowTie2-mapped SAM if you specify this format with \n"
" --input_type. Two steps: first apply BowTie2 and then feed MetaPhlAn2 with the obtained sam:\n"
"$ bowtie2 --sam-no-hd --sam-no-sq --no-unal --very-sensitive -S metagenome.sam -x /usr/share/metaphlan2/metaphlan_databases/mpa_v25_CHOCOPhlAn_201901 -U metagenome.fastq\n"
"$ metaphlan2.py metagenome.sam --input_type sam -o profiled_metagenome.txt\n\n"
"* We can also natively handle paired-end metagenomes, and, more generally, metagenomes stored in \n"
" multiple files (but you need to specify the --bowtie2out parameter):\n"
"$ metaphlan2.py metagenome_1.fastq,metagenome_2.fastq --bowtie2out metagenome.bowtie2.bz2 --nproc 5 --input_type fastq\n\n"
"\n------------------------------------------------------------------- \n \n\n"
"\n========== Marker level analysis ============================ \n\n"
"MetaPhlAn 2 introduces the capability of charachterizing organisms at the strain level using non\n"
"aggregated marker information. Such capability comes with several slightly different flavours and \n"
"are a way to perform strain tracking and comparison across multiple samples.\n"
"Usually, MetaPhlAn 2 is first ran with the default -t to profile the species present in\n"
"the community, and then a strain-level profiling can be performed to zoom-in into specific species\n"
"of interest. This operation can be performed quickly as it exploits the --bowtie2out intermediate \n"
"file saved during the execution of the default analysis type.\n\n"
"* The following command will output the abundance of each marker with a RPK (reads per kilo-base) \n"
" higher 0.0. (we are assuming that metagenome_outfmt.bz2 has been generated before as \n"
" shown above).\n"
"$ metaphlan2.py -t marker_ab_table metagenome_outfmt.bz2 --input_type bowtie2out -o marker_abundance_table.txt\n"
" The obtained RPK can be optionally normalized by the total number of reads in the metagenome \n"
" to guarantee fair comparisons of abundances across samples. The number of reads in the metagenome\n"
" needs to be passed with the '--nreads' argument\n\n"
"* The list of markers present in the sample can be obtained with '-t marker_pres_table'\n"
"$ metaphlan2.py -t marker_pres_table metagenome_outfmt.bz2 --input_type bowtie2out -o marker_abundance_table.txt\n"
" The --pres_th argument (default 1.0) set the minimum RPK value to consider a marker present\n\n"
"* The list '-t clade_profiles' analysis type reports the same information of '-t marker_ab_table'\n"
" but the markers are reported on a clade-by-clade basis.\n"
"$ metaphlan2.py -t clade_profiles metagenome_outfmt.bz2 --input_type bowtie2out -o marker_abundance_table.txt\n\n"
"* Finally, to obtain all markers present for a specific clade and all its subclades, the \n"
" '-t clade_specific_strain_tracker' should be used. For example, the following command\n"
" is reporting the presence/absence of the markers for the B. fragulis species and its strains\n"
" the optional argument --min_ab specifies the minimum clade abundance for reporting the markers\n\n"
"$ metaphlan2.py -t clade_specific_strain_tracker --clade s__Bacteroides_fragilis metagenome_outfmt.bz2 --input_type bowtie2out -o marker_abundance_table.txt\n"
"\n------------------------------------------------------------------- \n\n"
"",
formatter_class=ap.RawTextHelpFormatter,
add_help=False )
arg = p.add_argument
arg( 'inp', metavar='INPUT_FILE', type=str, nargs='?', default=None, help=
"the input file can be:\n"
"* a fastq file containing metagenomic reads\n"
"OR\n"
"* a BowTie2 produced SAM file. \n"
"OR\n"
"* an intermediary mapping file of the metagenome generated by a previous MetaPhlAn run \n"
"If the input file is missing, the script assumes that the input is provided using the standard \n"
"input, or named pipes.\n"
"IMPORTANT: the type of input needs to be specified with --input_type" )
arg( 'output', metavar='OUTPUT_FILE', type=str, nargs='?', default=None,
help= "the tab-separated output file of the predicted taxon relative abundances \n"
"[stdout if not present]")
g = p.add_argument_group('Required arguments')
arg = g.add_argument
input_type_choices = ['fastq','fasta','multifasta','multifastq','bowtie2out','sam']
arg( '--input_type', choices=input_type_choices, required = '--install' not in args, help =
"set whether the input is the multifasta file of metagenomic reads or \n"
"the SAM file of the mapping of the reads against the MetaPhlAn db.\n"
"[default 'automatic', i.e. the script will try to guess the input format]\n" )
g = p.add_argument_group('Mapping arguments')
arg = g.add_argument
arg('--mpa_pkl', type=str, default=None,
help="The metadata pickled MetaPhlAn file [deprecated]")
arg('--force', action='store_true', help="Force profiling of the input file by removing the bowtie2out file")
arg('--bowtie2db', metavar="METAPHLAN_BOWTIE2_DB", type=str, default=DEFAULT_DB_FOLDER,
help=("Folder containing the MetaPhlAn database. Used if "
"--input_type is fastq, fasta, multifasta, or multifastq [default "+DEFAULT_DB_FOLDER+"]\n"))
INDEX = 'latest'
arg('-x', '--index', type=str, default=INDEX,
help=("Specify the id of the database version to use. "
"If \"latest\", MetaPhlAn2 will get the latest version. If the database\n"
"files are not found on the local MetaPhlAn2 installation they\n"
"will be automatically downloaded [default "+INDEX+"]\n"))
bt2ps = ['sensitive', 'very-sensitive', 'sensitive-local', 'very-sensitive-local']
arg('--bt2_ps', metavar="BowTie2 presets", default='very-sensitive',
choices=bt2ps, help="Presets options for BowTie2 (applied only when a "
"multifasta file is provided)\n"
"The choices enabled in MetaPhlAn are:\n"
" * sensitive\n"
" * very-sensitive\n"
" * sensitive-local\n"
" * very-sensitive-local\n"
"[default very-sensitive]\n")
arg('--bowtie2_exe', type=str, default=None,
help='Full path and name of the BowTie2 executable. This option allows'
'MetaPhlAn to reach the executable even when it is not in the '
'system PATH or the system PATH is unreachable')
arg('--bowtie2_build', type=str, default='bowtie2-build',
help="Full path to the bowtie2-build command to use, deafult assumes "
"that 'bowtie2-build is present in the system path")
arg('--bowtie2out', metavar="FILE_NAME", type=str, default=None,
help="The file for saving the output of BowTie2")
arg('--min_mapq_val', type=int, default="5",
help="Minimum mapping quality value (MAPQ)")
arg('--no_map', action='store_true',
help="Avoid storing the --bowtie2out map file")
arg('--tmp_dir', metavar="", default=None, type=str,
help="The folder used to store temporary files [default is the OS "
"dependent tmp dir]")
g = p.add_argument_group('Post-mapping arguments')
arg = g.add_argument
stat_choices = ['avg_g','avg_l','tavg_g','tavg_l','wavg_g','wavg_l','med']
arg( '--tax_lev', metavar='TAXONOMIC_LEVEL', type=str,
choices='a'+tax_units, default='a', help =
"The taxonomic level for the relative abundance output:\n"
"'a' : all taxonomic levels\n"
"'k' : kingdoms\n"
"'p' : phyla only\n"
"'c' : classes only\n"
"'o' : orders only\n"
"'f' : families only\n"
"'g' : genera only\n"
"'s' : species only\n"
"[default 'a']" )
arg( '--min_cu_len', metavar="", default="2000", type=int, help =
"minimum total nucleotide length for the markers in a clade for\n"
"estimating the abundance without considering sub-clade abundances\n"
"[default 2000]\n" )
arg( '--min_alignment_len', metavar="", default=None, type=int, help =
"The sam records for aligned reads with the longest subalignment\n"
"length smaller than this threshold will be discarded.\n"
"[default None]\n" )
arg( '--ignore_eukaryotes', action='store_true', help=
"Do not profile eukaryotic organisms" )
arg( '--ignore_bacteria', action='store_true', help=
"Do not profile bacterial organisms" )
arg( '--ignore_archaea', action='store_true', help=
"Do not profile archeal organisms" )
arg( '--stat_q', metavar="", type = float, default=0.1, help =
"Quantile value for the robust average\n"
"[default 0.1]" )
arg( '--perc_nonzero', metavar="", type = float, default=0.33, help =
"Percentage of markers with a non zero relative abundance for misidentify a species\n"
"[default 0.33]" )
arg( '--ignore_markers', type=str, default = None, help =
"File containing a list of markers to ignore. \n")
arg( '--avoid_disqm', action="store_true", help =
"Deactivate the procedure of disambiguating the quasi-markers based on the \n"
"marker abundance pattern found in the sample. It is generally recommended \n"
"to keep the disambiguation procedure in order to minimize false positives\n")
arg( '--stat', metavar="", choices=stat_choices, default="tavg_g", type=str, help =
"EXPERIMENTAL! Statistical approach for converting marker abundances into clade abundances\n"
"'avg_g' : clade global (i.e. normalizing all markers together) average\n"
"'avg_l' : average of length-normalized marker counts\n"
"'tavg_g' : truncated clade global average at --stat_q quantile\n"
"'tavg_l' : trunated average of length-normalized marker counts (at --stat_q)\n"
"'wavg_g' : winsorized clade global average (at --stat_q)\n"
"'wavg_l' : winsorized average of length-normalized marker counts (at --stat_q)\n"
"'med' : median of length-normalized marker counts\n"
"[default tavg_g]" )
arg = p.add_argument
g = p.add_argument_group('Additional analysis types and arguments')
arg = g.add_argument
analysis_types = ['rel_ab', 'rel_ab_w_read_stats', 'reads_map', 'clade_profiles', 'marker_ab_table', 'marker_counts', 'marker_pres_table', 'clade_specific_strain_tracker']
arg( '-t', metavar='ANALYSIS TYPE', type=str, choices = analysis_types,
default='rel_ab', help =
"Type of analysis to perform: \n"
" * rel_ab: profiling a metagenomes in terms of relative abundances\n"
" * rel_ab_w_read_stats: profiling a metagenomes in terms of relative abundances and estimate the number of reads coming from each clade.\n"
" * reads_map: mapping from reads to clades (only reads hitting a marker)\n"
" * clade_profiles: normalized marker counts for clades with at least a non-null marker\n"
" * marker_ab_table: normalized marker counts (only when > 0.0 and normalized by metagenome size if --nreads is specified)\n"
" * marker_counts: non-normalized marker counts [use with extreme caution]\n"
" * marker_pres_table: list of markers present in the sample (threshold at 1.0 if not differently specified with --pres_th\n"
"[default 'rel_ab']" )
arg( '--nreads', metavar="NUMBER_OF_READS", type=int, default = None, help =
"The total number of reads in the original metagenome. It is used only when \n"
"-t marker_table is specified for normalizing the length-normalized counts \n"
"with the metagenome size as well. No normalization applied if --nreads is not \n"
"specified" )
arg( '--pres_th', metavar="PRESENCE_THRESHOLD", type=int, default = 1.0, help =
'Threshold for calling a marker present by the -t marker_pres_table option' )
arg( '--clade', metavar="", default=None, type=str, help =
"The clade for clade_specific_strain_tracker analysis\n" )
arg( '--min_ab', metavar="", default=0.1, type=float, help =
"The minimum percentage abundace for the clade in the clade_specific_strain_tracker analysis\n" )
g = p.add_argument_group('Output arguments')
arg = g.add_argument
arg( '-o', '--output_file', metavar="output file", type=str, default=None, help =
"The output file (if not specified as positional argument)\n")
arg('--sample_id_key', metavar="name", type=str, default="#SampleID",
help =("Specify the sample ID key for this analysis."
" Defaults to '#SampleID'."))
arg('--sample_id', metavar="value", type=str,
default="Metaphlan2_Analysis",
help =("Specify the sample ID for this analysis."
" Defaults to 'Metaphlan2_Analysis'."))
arg( '-s', '--samout', metavar="sam_output_file",
type=str, default=None, help="The sam output file\n")
arg( '--legacy-output', action='store_true', help="Old two columns output\n")
arg( '--CAMI_format_output', action='store_true', help="Report the profiling using the CAMI output format\n")
arg( '--unknown_estimation', action='store_true', help="Ignore estimation of reads mapping to unkwnown clades\n")
#*************************************************************
#* Parameters related to biom file generation *
#*************************************************************
arg( '--biom', '--biom_output_file', metavar="biom_output", type=str, default=None, help =
"If requesting biom file output: The name of the output file in biom format \n")
arg( '--mdelim', '--metadata_delimiter_char', metavar="mdelim", type=str, default="|", help =
"Delimiter for bug metadata: - defaults to pipe. e.g. the pipe in k__Bacteria|p__Proteobacteria \n")
#*************************************************************
#* End parameters related to biom file generation *
#*************************************************************
g = p.add_argument_group('Other arguments')
arg = g.add_argument
arg('--nproc', metavar="N", type=int, default=4,
help="The number of CPUs to use for parallelizing the mapping [default 4]")
arg('--install', action='store_true',
help="Only checks if the MetaPhlAn2 DB is installed and installs it if not. All other parameters are ignored.")
arg('--force_download', action='store_true',
help="Force the re-download of the latest MetaPhlAn2 database.")
arg('--read_min_len', type=int, default=70,
help="Specify the minimum length of the reads to be considered when parsing the input file with "
"'read_fastx.py' script, default value is 70")
arg('-v', '--version', action='version',
version="MetaPhlAn version {} ({})".format(__version__, __date__),
help="Prints the current MetaPhlAn version and exit")
arg("-h", "--help", action="help", help="show this help message and exit")
return vars(p.parse_args())
def byte_to_megabyte(byte):
"""
Convert byte value to megabyte
"""
return byte / (1024.0**2)
class ReportHook():
def __init__(self):
self.start_time = time.time()
def report(self, blocknum, block_size, total_size):
"""
Print download progress message
"""
if blocknum == 0:
self.start_time = time.time()
if total_size > 0:
sys.stderr.write("Downloading file of size: {:.2f} MB\n"
.format(byte_to_megabyte(total_size)))
else:
total_downloaded = blocknum * block_size
status = "{:3.2f} MB ".format(byte_to_megabyte(total_downloaded))
if total_size > 0:
percent_downloaded = total_downloaded * 100.0 / total_size
# use carriage return plus sys.stderr to overwrite stderr
download_rate = total_downloaded / (time.time() - self.start_time)
estimated_time = (total_size - total_downloaded) / download_rate
estimated_minutes = int(estimated_time / 60.0)
estimated_seconds = estimated_time - estimated_minutes * 60.0
status += ("{:3.2f} % {:5.2f} MB/sec {:2.0f} min {:2.0f} sec "
.format(percent_downloaded,
byte_to_megabyte(download_rate),
estimated_minutes, estimated_seconds))
status += " \r"
sys.stderr.write(status)
def download(url, download_file, force=False):
"""
Download a file from a url
"""
if not os.path.isfile(download_file) or force:
try:
sys.stderr.write("\nDownloading " + url + "\n")
file, headers = urlretrieve(url, download_file,
reporthook=ReportHook().report)
except EnvironmentError:
sys.stderr.write("\nWarning: Unable to download " + url + "\n")
else:
sys.stderr.write("\nFile {} already present!\n".format(download_file))
def download_unpack_tar(url, download_file_name, folder, bowtie2_build, nproc):
"""
Download the url to the file and decompress into the folder
"""
tar_file = os.path.join(folder, download_file_name + ".tar")
url_tar_file = os.path.join(url, download_file_name + ".tar")
download(url_tar_file, tar_file)
# download MD5 checksum
md5_file = os.path.join(folder, download_file_name + ".md5")
url_md5_file = os.path.join(url, download_file_name + ".md5")
download(url_md5_file, md5_file)
md5_md5 = None
md5_tar = None
if os.path.isfile(md5_file):
with open(md5_file) as f:
for row in f:
md5_md5 = row.strip().split(' ')[0]
else:
sys.stderr.write('File "{}" not found!\n'.format(md5_file))
# compute MD5 of .tar.bz2
if os.path.isfile(tar_file):
hash_md5 = hashlib.md5()
with open(tar_file, "rb") as f:
for chunk in iter(lambda: f.read(4096), b""):
hash_md5.update(chunk)
md5_tar = hash_md5.hexdigest()[:32]
else:
sys.stderr.write('File "{}" not found!\n'.format(tar_file))
if (md5_tar is None) or (md5_md5 is None):
sys.exit("MD5 checksums not found, something went wrong!")
# compare checksums
if md5_tar != md5_md5:
sys.exit("MD5 checksums do not correspond! If this happens again, you should remove the database files and "
"rerun MetaPhlAn2 so they are re-downloaded")
# untar
try:
tarfile_handle = tarfile.open(tar_file)
tarfile_handle.extractall(path=folder)
tarfile_handle.close()
except EnvironmentError:
sys.stderr.write("Warning: Unable to extract {}.\n".format(tar_file))
# uncompress sequences
bz2_file = os.path.join(folder, download_file_name + ".fna.bz2")
fna_file = os.path.join(folder, download_file_name + ".fna")
if not os.path.isfile(fna_file):
sys.stderr.write('\n\nDecompressing {} into {}\n'.format(bz2_file, fna_file))
with open(fna_file, 'wb') as fna_h, \
bz2.BZ2File(bz2_file, 'rb') as bz2_h:
for data in iter(lambda: bz2_h.read(100 * 1024), b''):
fna_h.write(data)
# build bowtie2 indexes
if not glob(os.path.join(folder, download_file_name + "*.bt2")):
bt2_base = os.path.join(folder, download_file_name)
bt2_cmd = [bowtie2_build, '--quiet']
if nproc > 1:
bt2_build_output = subp.check_output([bowtie2_build, '--usage'], stderr=subp.STDOUT)
if 'threads' in str(bt2_build_output):
bt2_cmd += ['--threads', str(nproc)]
bt2_cmd += ['-f', fna_file, bt2_base]
sys.stderr.write('\nBuilding Bowtie2 indexes\n')
try:
subp.check_call(bt2_cmd)
except Exception as e:
sys.stderr.write("Fatal error running '{}'\nError message: '{}'\n\n".format(' '.join(bt2_cmd), e))
sys.exit(1)
for bt2 in glob(os.path.join(folder, download_file_name + "*.bt2")):
os.chmod(bt2, stat.S_IRUSR | stat.S_IWUSR | stat.S_IRGRP | stat.S_IWGRP | stat.S_IROTH) # change permissions to 664
sys.stderr.write('Removing uncompress database {}\n'.format(fna_file))
os.remove(fna_file)
def resolve_latest_database(bowtie2_db, force=False):
if os.path.exists(os.path.join(bowtie2_db,'mpa_latest')):
ctime_latest_db = int(os.path.getctime(os.path.join(bowtie2_db,'mpa_latest')))
if int(time.time()) - ctime_latest_db > 2419200: #1 month in epoch
os.remove(os.path.join(bowtie2_db,'mpa_latest'))
download(DATABASE_DOWNLOAD+'mpa_latest', os.path.join(bowtie2_db,'mpa_latest'), force=True)
if not os.path.exists(os.path.join(bowtie2_db,'mpa_latest') or force):
download(DATABASE_DOWNLOAD+'mpa_latest', os.path.join(bowtie2_db,'mpa_latest'))
with open(os.path.join(bowtie2_db,'mpa_latest')) as mpa_latest:
latest_db_version = [line.strip() for line in mpa_latest if not line.startswith('#')]
return ''.join(latest_db_version)
def check_and_install_database(index, bowtie2_db, bowtie2_build, nproc, force_redownload_latest):
# Create the folder if it does not already exist
if not os.path.isdir(bowtie2_db):
try:
os.makedirs(bowtie2_db)
except EnvironmentError:
sys.exit("ERROR: Unable to create folder for database install: " + bowtie2_db)
# Check the directory permissions
if not os.access(bowtie2_db, os.W_OK):
sys.exit("ERROR: The directory is not writeable: " + bowtie2_db + ". "
"Please modify the permissions.")
""" Check if the database is installed, if not download and install """
if index == 'latest':
index = resolve_latest_database(bowtie2_db, force_redownload_latest)
if len(glob(os.path.join(bowtie2_db, "*{}*".format(index)))) >= 7:
return index
# download the tar archive and decompress
sys.stderr.write("\nDownloading MetaPhlAn2 database\nPlease note due to "
"the size this might take a few minutes\n")
download_unpack_tar(DATABASE_DOWNLOAD, index, bowtie2_db, bowtie2_build, nproc)
sys.stderr.write("\nDownload complete\n")
return index
def set_mapping_arguments(index, bowtie2_db):
mpa_pkl = 'mpa_pkl'
bowtie2db = 'bowtie2db'
if os.path.isfile(os.path.join(bowtie2_db, "{}.pkl".format(index))):
mpa_pkl = os.path.join(bowtie2_db, "{}.pkl".format(index))
if glob(os.path.join(bowtie2_db, "{}*.bt2".format(index))):
bowtie2db = os.path.join(bowtie2_db, "{}".format(index))
return (mpa_pkl, bowtie2db)
def run_bowtie2(fna_in, outfmt6_out, bowtie2_db, preset, nproc, min_mapq_val, file_format="multifasta",
exe=None, samout=None, min_alignment_len=None, read_min_len=0):
# checking read_fastx.py
read_fastx = "read_fastx.py"
try:
subp.check_call([read_fastx, "-h"], stdout=DEVNULL, stderr=DEVNULL)
except Exception as e:
try:
read_fastx = os.path.join(os.path.join(os.path.dirname(__file__), "utils"), read_fastx)
subp.check_call([read_fastx, "-h"], stdout=DEVNULL, stderr=DEVNULL)
except Exception as e:
sys.stderr.write("OSError: fatal error running '{}'. Is it in the system path?\n".format(read_fastx))
sys.exit(1)
# checking bowtie2
try:
subp.check_call([exe if exe else 'bowtie2', "-h"], stdout=DEVNULL)
except Exception as e:
sys.stderr.write('OSError: "{}"\nFatal error running BowTie2. Is BowTie2 in the system path?\n'.format(e))
sys.exit(1)
try:
if fna_in:
readin = subp.Popen([read_fastx, '-l', str(read_min_len), fna_in], stdout=subp.PIPE, stderr=subp.PIPE)
else:
readin = subp.Popen([read_fastx, '-l', str(read_min_len)], stdin=sys.stdin, stdout=subp.PIPE, stderr=subp.PIPE)
bowtie2_cmd = [exe if exe else 'bowtie2', "--quiet", "--no-unal", "--{}".format(preset),
"-S", "-", "-x", bowtie2_db]
if int(nproc) > 1:
bowtie2_cmd += ["-p", str(nproc)]
bowtie2_cmd += ["-U", "-"] # if not stat.S_ISFIFO(os.stat(fna_in).st_mode) else []
if file_format == "multifasta":
bowtie2_cmd += ["-f"]
p = subp.Popen(bowtie2_cmd, stdout=subp.PIPE, stdin=readin.stdout)
readin.stdout.close()
lmybytes, outf = (mybytes, bz2.BZ2File(outfmt6_out, "w")) if outfmt6_out.endswith(".bz2") else (str, open(outfmt6_out, "w"))
try:
if samout:
if samout[-4:] == '.bz2':
sam_file = bz2.BZ2File(samout, 'w')
else:
sam_file = open(samout, 'wb')
except IOError as e:
sys.stderr.write('IOError: "{}"\nUnable to open sam output file.\n'.format(e))
sys.exit(1)
for line in p.stdout:
if samout:
sam_file.write(line)
o = read_and_split_line(line)
if not o[0].startswith('@'):
if not o[2].endswith('*'):
if (hex(int(o[1]) & 0x100) == '0x0'): #no secondary
if (int(o[4]) > min_mapq_val ): # filter low mapq reads
if ((min_alignment_len is None) or
(max([int(x.strip('M')) for x in re.findall(r'(\d*M)', o[5]) if x]) >= min_alignment_len)):
outf.write(lmybytes("\t".join([o[0], o[2]]) + "\n"))
if samout:
sam_file.close()
p.communicate()
n_metagenome_reads = ''.join(read_and_split_line(readin.stderr.readline()))
if not len(n_metagenome_reads):
sys.stderr.write('Fatal error running MetaPhlAn2. Total metagenome size was not estimated.\nPlease update read_fastx.py to the latest version.\n')
sys.exit(1)
outf.write(lmybytes('#nreads\t{}'.format(n_metagenome_reads)))
outf.close()
except OSError as e:
sys.stderr.write('OSError: "{}"\nFatal error running BowTie2.\n'.format(e))
sys.exit(1)
except ValueError as e:
sys.stderr.write('ValueError: "{}"\nFatal error running BowTie2.\n'.format(e))
sys.exit(1)
except IOError as e:
sys.stderr.write('IOError: "{}"\nFatal error running BowTie2.\n'.format(e))
sys.exit(1)
if p.returncode == 13:
sys.stderr.write("Permission Denied Error: fatal error running BowTie2."
"Is the BowTie2 file in the path with execution and read permissions?\n")
sys.exit(1)
elif p.returncode != 0:
sys.stderr.write("Error while running bowtie2.\n")
sys.exit(1)
class TaxClade:
min_cu_len = -1
markers2lens = None
stat = None
perc_nonzero = None
quantile = None
avoid_disqm = False
def __init__( self, name, tax_id, uncl = False):
self.children, self.markers2nreads = {}, {}
self.name, self.father = name, None
self.uncl, self.subcl_uncl = uncl, False
self.abundance, self.uncl_abundance = None, 0
self.nreads, self.uncl_nreads = 0, 0
self.tax_id = tax_id
def add_child( self, name, tax_id ):
new_clade = TaxClade( name, tax_id )
self.children[name] = new_clade
new_clade.father = self
return new_clade
def get_terminals( self ):
terms = []
if not self.children:
return [self]
for c in self.children.values():
terms += c.get_terminals()
return terms
def get_full_taxids( self ):
if self.tax_id:
fullname = [self.tax_id]
cl = self.father
while cl:
fullname = [cl.tax_id] + fullname
cl = cl.father
return "|".join(fullname[1:])
return ""
def get_full_name( self ):
fullname = [self.name]
cl = self.father
while cl:
fullname = [cl.name] + fullname
cl = cl.father
return "|".join(fullname[1:])
def get_normalized_counts( self ):
return [(m,float(n)*1000.0/self.markers2lens[m])
for m,n in self.markers2nreads.items()]
def compute_mapped_reads( self ):
if self.name.startswith('s__'):
return self.nreads
for c in self.children.values():
self.nreads += c.compute_mapped_reads()
return self.nreads
def compute_abundance( self ):
if self.abundance is not None: return self.abundance
sum_ab = sum([c.compute_abundance() for c in self.children.values()])
# rat_nreads = sorted([(self.markers2lens[marker], n_reads)
# for marker,n_reads in self.markers2nreads.items()],
# key = lambda x: x[1])
rat_nreads, removed = [], []
for marker, n_reads in sorted(self.markers2nreads.items(),key=lambda x:x[0]):
misidentified = False
if not self.avoid_disqm:
for ext in self.markers2exts[marker]:
ext_clade = self.taxa2clades[ext]
m2nr = ext_clade.markers2nreads
tocladetmp = ext_clade
while len(tocladetmp.children) == 1:
tocladetmp = list(tocladetmp.children.values())[0]
m2nr = tocladetmp.markers2nreads
nonzeros = sum([v>0 for v in m2nr.values()])
if len(m2nr):
if float(nonzeros) / len(m2nr) > self.perc_nonzero:
misidentified = True
removed.append( (self.markers2lens[marker],n_reads) )
break
if not misidentified:
rat_nreads.append( (self.markers2lens[marker],n_reads) )
if not self.avoid_disqm and len(removed):
n_rat_nreads = float(len(rat_nreads))
n_removed = float(len(removed))
n_tot = n_rat_nreads + n_removed
n_ripr = 10
if len(self.get_terminals()) < 2:
n_ripr = 0
if "k__Viruses" in self.get_full_name():
n_ripr = 0
if n_rat_nreads < n_ripr and n_tot > n_rat_nreads:
rat_nreads += removed[:n_ripr-int(n_rat_nreads)]
rat_nreads = sorted(rat_nreads, key = lambda x: x[1])
rat_v,nreads_v = zip(*rat_nreads) if rat_nreads else ([],[])
rat, nrawreads, loc_ab = float(sum(rat_v)) or -1.0, sum(nreads_v), 0.0
quant = int(self.quantile*len(rat_nreads))
ql,qr,qn = (quant,-quant,quant) if quant else (None,None,0)
if self.name[0] == 't' and (len(self.father.children) > 1 or "_sp" in self.father.name or "k__Viruses" in self.get_full_name()):
non_zeros = float(len([n for r,n in rat_nreads if n > 0]))
nreads = float(len(rat_nreads))
if nreads == 0.0 or non_zeros / nreads < 0.7:
self.abundance = 0.0
return 0.0
if rat < 0.0:
pass
elif self.stat == 'avg_g' or (not qn and self.stat in ['wavg_g','tavg_g']):
loc_ab = nrawreads / rat if rat >= 0 else 0.0
elif self.stat == 'avg_l' or (not qn and self.stat in ['wavg_l','tavg_l']):
loc_ab = np.mean([float(n)/r for r,n in rat_nreads])
elif self.stat == 'tavg_g':
wnreads = sorted([(float(n)/r,r,n) for r,n in rat_nreads], key=lambda x:x[0])
den,num = zip(*[v[1:] for v in wnreads[ql:qr]])
loc_ab = float(sum(num))/float(sum(den)) if any(den) else 0.0
elif self.stat == 'tavg_l':
loc_ab = np.mean(sorted([float(n)/r for r,n in rat_nreads])[ql:qr])
elif self.stat == 'wavg_g':
vmin, vmax = nreads_v[ql], nreads_v[qr]
wnreads = [vmin]*qn+list(nreads_v[ql:qr])+[vmax]*qn
loc_ab = float(sum(wnreads)) / rat
elif self.stat == 'wavg_l':
wnreads = sorted([float(n)/r for r,n in rat_nreads])
vmin, vmax = wnreads[ql], wnreads[qr]
wnreads = [vmin]*qn+list(wnreads[ql:qr])+[vmax]*qn
loc_ab = np.mean(wnreads)
elif self.stat == 'med':
loc_ab = np.median(sorted([float(n)/r for r,n in rat_nreads])[ql:qr])
self.abundance = loc_ab
if rat < self.min_cu_len and self.children:
self.abundance = sum_ab
elif loc_ab < sum_ab:
self.abundance = sum_ab
if self.abundance > sum_ab and self.children: # *1.1??
self.uncl_abundance = self.abundance - sum_ab
self.subcl_uncl = not self.children and self.name[0] not in tax_units[-2:]
return self.abundance
def get_all_abundances( self ):
ret = [(self.name, self.tax_id, self.abundance)]
if self.uncl_abundance > 0.0:
lchild = list(self.children.values())[0].name[:3]
ret += [(lchild+self.name[3:]+"_unclassified", "", self.uncl_abundance)]
if self.subcl_uncl and self.name[0] != tax_units[-2]:
cind = tax_units.index( self.name[0] )
ret += [( tax_units[cind+1]+self.name[1:]+"_unclassified","",
self.abundance)]
for c in self.children.values():
ret += c.get_all_abundances()
return ret
class TaxTree:
def __init__( self, mpa, markers_to_ignore = None ): #, min_cu_len ):
self.root = TaxClade( "root", 0)
self.all_clades, self.markers2lens, self.markers2clades, self.taxa2clades, self.markers2exts = {}, {}, {}, {}, {}
TaxClade.markers2lens = self.markers2lens
TaxClade.markers2exts = self.markers2exts
TaxClade.taxa2clades = self.taxa2clades
for clade, value in mpa['taxonomy'].items():
clade = clade.strip().split("|")
if isinstance(value,tuple):
taxids, lenc = value
taxids = taxids.strip().split("|")
if isinstance(value,int):
lenc = value
taxids = None
father = self.root
for i in range(len(clade)):
clade_lev = clade[i]
clade_taxid = taxids[i] if i < 7 and taxids is not None else None
if not clade_lev in father.children:
father.add_child(clade_lev, tax_id=clade_taxid)
self.all_clades[clade_lev] = father.children[clade_lev]
if clade_lev[0] == "t":
self.taxa2clades[clade_lev[3:]] = father
father = father.children[clade_lev]
if clade_lev[0] == "t":
father.glen = lenc
def add_lens( node ):
if not node.children:
return node.glen
lens = []
for c in node.children.values():
lens.append( add_lens( c ) )
node.glen = min(np.mean(lens), np.median(lens))
return node.glen
add_lens(self.root)
# for k,p in mpa_pkl['markers'].items():
for k, p in mpa['markers'].items():
if k in markers_to_ignore:
continue
self.markers2lens[k] = p['len']
self.markers2clades[k] = p['clade']
self.add_reads(k, 0)
self.markers2exts[k] = p['ext']
def set_min_cu_len( self, min_cu_len ):
TaxClade.min_cu_len = min_cu_len
def set_stat( self, stat, quantile, perc_nonzero, avoid_disqm = False):
TaxClade.stat = stat
TaxClade.perc_nonzero = perc_nonzero
TaxClade.quantile = quantile
TaxClade.avoid_disqm = avoid_disqm
def add_reads( self, marker, n,
ignore_eukaryotes = False,
ignore_bacteria = False, ignore_archaea = False ):
clade = self.markers2clades[marker]
cl = self.all_clades[clade]
if ignore_eukaryotes or ignore_bacteria or ignore_archaea:
cn = cl.get_full_name()
if ignore_eukaryotes and cn.startswith("k__Eukaryota"):
return (None, None)
if ignore_archaea and cn.startswith("k__Archaea"):
return (None, None)
if ignore_bacteria and cn.startswith("k__Bacteria"):
return (None, None)
# while len(cl.children) == 1:
# cl = list(cl.children.values())[0]
cl.markers2nreads[marker] = n
return (cl.get_full_name(), cl.get_full_taxids(), )
def markers2counts( self ):
m2c = {}
for k,v in self.all_clades.items():
for m,c in v.markers2nreads.items():
m2c[m] = c
return m2c
def clade_profiles( self, tax_lev, get_all = False ):
cl2pr = {}
for k,v in self.all_clades.items():
if tax_lev and not k.startswith(tax_lev):
continue
prof = v.get_normalized_counts()
if not get_all and ( len(prof) < 1 or not sum([p[1] for p in prof]) > 0.0 ):
continue
cl2pr[v.get_full_name()] = prof
return cl2pr
def relative_abundances( self, tax_lev ):
clade2abundance_n = dict([(tax_label, clade) for tax_label, clade in self.all_clades.items()
if tax_label.startswith("k__") and not clade.uncl])
clade2abundance, clade2est_nreads, tot_ab, tot_reads = {}, {}, 0.0, 0
for tax_label, clade in clade2abundance_n.items():
tot_ab += clade.compute_abundance()
for tax_label, clade in clade2abundance_n.items():
for clade_label, tax_id, abundance in sorted(clade.get_all_abundances(), key=lambda pars:pars[0]):
if clade_label[:3] != 't__':
if not tax_lev:
if clade_label not in self.all_clades:
to = tax_units.index(clade_label[0])
t = tax_units[to-1]
clade_label = t + clade_label.split("_unclassified")[0][1:]
tax_id = self.all_clades[clade_label].get_full_taxids()
clade_label = self.all_clades[clade_label].get_full_name()
spl = clade_label.split("|")
clade_label = "|".join(spl+[tax_units[to]+spl[-1][1:]+"_unclassified"])
glen = self.all_clades[spl[-1]].glen
else:
glen = self.all_clades[clade_label].glen
tax_id = self.all_clades[clade_label].get_full_taxids()
if 's__' in clade_label and abundance > 0:
self.all_clades[clade_label].nreads = int(np.floor(abundance*glen))
clade_label = self.all_clades[clade_label].get_full_name()
elif not clade_label.startswith(tax_lev):
if clade_label in self.all_clades:
glen = self.all_clades[clade_label].glen
else:
glen = 1.0
continue
clade2abundance[(clade_label, tax_id)] = abundance
for tax_label, clade in clade2abundance_n.items():
tot_reads += clade.compute_mapped_reads()
for clade_label, clade in self.all_clades.items():
nreads = clade.nreads
clade_label = clade.get_full_name()
tax_id = clade.get_full_taxids()
clade2est_nreads[(clade_label, tax_id)] = nreads
ret_d = dict([( tax, float(abundance) / tot_ab if tot_ab else 0.0) for tax, abundance in clade2abundance.items()])
ret_r = dict([( tax, (abundance, clade2est_nreads[tax] )) for tax, abundance in clade2abundance.items() if tax in clade2est_nreads])
if tax_lev:
ret_d[("UNKNOWN", '-1')] = 1.0 - sum(ret_d.values())
return ret_d, ret_r, tot_reads
def map2bbh(mapping_f, min_mapq_val, input_type='bowtie2out', min_alignment_len=None):
if not mapping_f:
ras, ras_line, inpf = plain_read_and_split, plain_read_and_split_line, sys.stdin
else:
if mapping_f.endswith(".bz2"):
ras, ras_line, inpf = read_and_split, read_and_split_line, bz2.BZ2File(mapping_f, "r")
else:
ras, ras_line, inpf = plain_read_and_split, plain_read_and_split_line, open(mapping_f)
reads2markers = {}
n_metagenoges_reads = None
if input_type == 'bowtie2out':
for r, c in ras(inpf):
if r.startswith('#') and 'nreads' in r:
n_metagenoges_reads = int(c)
else:
reads2markers[r] = c
elif input_type == 'sam':
for line in inpf:
o = ras_line(line)
if ((o[0][0] != '@') and #no header
(o[2][-1] != '*') and # no unmapped reads
(hex(int(o[1]) & 0x100) == '0x0') and #no secondary
(int(o[4]) > min_mapq_val ) and # filter low mapq reads
( (min_alignment_len is None) or ( max(int(x.strip('M')) for x in re.findall(r'(\d*M)', o[5]) if x) >= min_alignment_len ) )
):
reads2markers[o[0]] = o[2]
inpf.close()
markers2reads = defdict(set)
for r, m in reads2markers.items():
markers2reads[m].add(r)
return (markers2reads, n_metagenoges_reads)
def maybe_generate_biom_file(tree, pars, abundance_predictions):
json_key = "MetaPhlAn2"
if not pars['biom']:
return None
if not abundance_predictions:
biom_table = biom.Table([], [], []) # create empty BIOM table
with open(pars['biom'], 'w') as outfile:
biom_table.to_json(json_key, direct_io=outfile)
return True
delimiter = "|" if len(pars['mdelim']) > 1 else pars['mdelim']
def istip(clade_name):
end_name = clade_name.split(delimiter)[-1]
return end_name.startswith("s__") or end_name.endswith("_unclassified")
def findclade(clade_name):
if clade_name.endswith('_unclassified'):
name = clade_name.split(delimiter)[-2]
else:
name = clade_name.split(delimiter)[-1]
return tree.all_clades[name]
def to_biomformat(clade_name):
return {'taxonomy': clade_name.split(delimiter)}
clades = iter((abundance, findclade(name))
for (name, taxid, abundance) in abundance_predictions if istip(name))
packed = iter(([abundance], clade.get_full_name(), clade.tax_id)
for (abundance, clade) in clades)
# unpack that tuple here to stay under 80 chars on a line
data, clade_names, clade_ids = zip(*packed)
# biom likes column vectors, so we give it an array like this:
# np.array([a],[b],[c])
data = np.array(data)
sample_ids = [pars['sample_id']]
table_id = 'MetaPhlAn2_Analysis'
#**********************************************************************************************
# Modification of Code : *
# Modified the code so instead of using the current clade IDs, which are numbers, we will *
# use the clade_names *
# Users reported the biom output is invalid and also the IDs were changing from run to *
# run. *
# George Weingart 05/22/2017 george.weingart@mail.com *
#**********************************************************************************************
if LooseVersion(biom.__version__) < LooseVersion("2.0.0"):
biom_table = biom.table.table_factory(
data,
sample_ids,
######## clade_ids, #Modified by George Weingart 5/22/2017 - We will use instead the clade_names
clade_names, #Modified by George Weingart 5/22/2017 - We will use instead the clade_names
sample_metadata = None,
observation_metadata = list(map(to_biomformat, clade_names)),
table_id = table_id,
constructor = biom.table.DenseOTUTable
)
with open(pars['biom'], 'w') as outfile:
json.dump( biom_table.getBiomFormatObject(json_key),
outfile )
else: # Below is the biom2 compatible code
biom_table = biom.table.Table(
data,
#clade_ids, #Modified by George Weingart 5/22/2017 - We will use instead the clade_names
clade_names, #Modified by George Weingart 5/22/2017 - We will use instead the clade_names
sample_ids,
sample_metadata = None,
observation_metadata = list(map(to_biomformat, clade_names)),
table_id = table_id,
input_is_dense = True
)
with open(pars['biom'], 'w') as outfile:
biom_table.to_json( json_key,
direct_io = outfile )
return True
def metaphlan2():
ranks2code = { 'k' : 'superkingdom', 'p' : 'phylum', 'c':'class',
'o' : 'order', 'f' : 'family', 'g' : 'genus', 's' : 'species'}
pars = read_params(sys.argv)
# check if the database is installed, if not then install
pars['index'] = check_and_install_database(pars['index'], pars['bowtie2db'], pars['bowtie2_build'], pars['nproc'], pars['force_download'])
if pars['install']:
sys.stderr.write('The database is installed\n')
return
# set correct map_pkl and bowtie2db variables
pars['mpa_pkl'], pars['bowtie2db'] = set_mapping_arguments(pars['index'], pars['bowtie2db'])
if (pars['bt2_ps'] in ["sensitive-local", "very-sensitive-local"]) and (pars['min_alignment_len'] is None):
pars['min_alignment_len'] = 100
sys.stderr.write('Warning! bt2_ps is set to local mode, and min_alignment_len is None, I automatically '
'set min_alignment_len to 100! If you do not like, rerun the command and set '
'min_alignment_len to a specific value.\n')
if pars['input_type'] == 'fastq':
pars['input_type'] = 'multifastq'
if pars['input_type'] == 'fasta':
pars['input_type'] = 'multifasta'
# check for the mpa_pkl file
if not os.path.isfile(pars['mpa_pkl']):
sys.stderr.write("Error: Unable to find the mpa_pkl file at: " + pars['mpa_pkl'] +
"Exiting...\n\n")
sys.exit(1)
if pars['ignore_markers']:
with open(pars['ignore_markers']) as ignv:
ignore_markers = set([l.strip() for l in ignv])
else:
ignore_markers = set()
no_map = False
if pars['input_type'] == 'multifasta' or pars['input_type'] == 'multifastq':
bow = pars['bowtie2db'] is not None
if not bow:
sys.stderr.write( "No MetaPhlAn BowTie2 database provided\n "
"[--bowtie2db and --index options]!\n"
"Exiting...\n\n" )
sys.exit(1)
if pars['no_map']:
pars['bowtie2out'] = tf.NamedTemporaryFile(dir=pars['tmp_dir']).name
no_map = True
else:
if bow and not pars['bowtie2out']:
if pars['inp'] and "," in pars['inp']:
sys.stderr.write("Error! --bowtie2out needs to be specified when multiple "
"fastq or fasta files (comma separated) are provided\n")
sys.exit(1)
fname = pars['inp']
if fname is None:
fname = "stdin_map"
elif stat.S_ISFIFO(os.stat(fname).st_mode):
fname = "fifo_map"
pars['bowtie2out'] = fname + ".bowtie2out.txt"
if os.path.exists( pars['bowtie2out'] ) and not pars['force']:
sys.stderr.write(
"BowTie2 output file detected: " + pars['bowtie2out'] + "\n"
"Please use it as input or remove it if you want to "
"re-perform the BowTie2 run.\n"
"Exiting...\n\n" )
sys.exit(1)
if pars['force']:
if os.path.exists(pars['bowtie2out']):
os.remove( pars['bowtie2out'] )
if bow and not all([os.path.exists(".".join([str(pars['bowtie2db']), p]))
for p in ["1.bt2", "2.bt2", "3.bt2", "4.bt2", "rev.1.bt2", "rev.2.bt2"]]):
sys.stderr.write("No MetaPhlAn BowTie2 database found (--index "
"option)!\nExpecting location {}\nExiting..."
.format(pars['bowtie2db']))
sys.exit(1)
if bow:
run_bowtie2(pars['inp'], pars['bowtie2out'], pars['bowtie2db'],
pars['bt2_ps'], pars['nproc'], file_format=pars['input_type'],
exe=pars['bowtie2_exe'], samout=pars['samout'],
min_alignment_len=pars['min_alignment_len'], read_min_len=pars['read_min_len'], min_mapq_val=pars['min_mapq_val'])
pars['input_type'] = 'bowtie2out'
pars['inp'] = pars['bowtie2out'] # !!!
with bz2.BZ2File( pars['mpa_pkl'], 'r' ) as a:
mpa_pkl = pickle.load( a )
tree = TaxTree( mpa_pkl, ignore_markers )
tree.set_min_cu_len( pars['min_cu_len'] )
tree.set_stat( pars['stat'], pars['stat_q'], pars['perc_nonzero'], pars['avoid_disqm'])
markers2reads, n_metagenome_reads = map2bbh(pars['inp'], pars['min_mapq_val'], pars['input_type'], pars['min_alignment_len'])
if no_map:
os.remove( pars['inp'] )
if not n_metagenome_reads and not pars['nreads']:
sys.stderr.write(
"Please provide the size of the metagenome using the "
"--nreads parameter when running MetaPhlAn2"
"\nExiting...\n\n" )
sys.exit(1)
map_out = []
for marker,reads in sorted(markers2reads.items(), key=lambda pars: pars[0]):
if marker not in tree.markers2lens:
continue
tax_seq, ids_seq = tree.add_reads( marker, len(reads),
ignore_eukaryotes = pars['ignore_eukaryotes'],
ignore_bacteria = pars['ignore_bacteria'],
ignore_archaea = pars['ignore_archaea'],
)
if tax_seq:
map_out +=["\t".join([r,tax_seq, ids_seq]) for r in sorted(reads)]
if pars['output'] is None and pars['output_file'] is not None:
pars['output'] = pars['output_file']
with (open(pars['output'],"w") if pars['output'] else sys.stdout) as outf:
if not pars['legacy_output']:
outf.write('#{}\n'.format(pars['index']))
outf.write('#{}\n'.format(' '.join(sys.argv)))
if not pars['CAMI_format_output']:
outf.write('\t'.join((pars["sample_id_key"], pars["sample_id"])) + '\n')
if pars['t'] == 'reads_map':
if not pars['legacy_output']:
outf.write('#read_id\tNCBI_taxlineage_str\tNCBI_taxlineage_ids\n')
outf.write( "\n".join( map_out ) + "\n" )
elif pars['t'] == 'rel_ab':
if pars['CAMI_format_output']:
outf.write("@SampleID:{}\n"
"@Version:0.10.0\n"
"@Ranks:superkingdom|phylum|class|order|family|genus|species|strain\n"
"@@TAXID\tRANK\tTAXPATH\tTAXPATHSN\tPERCENTAGE\n".format(pars["sample_id"],__version__))
elif not pars['legacy_output']:
outf.write('#clade_name\tNCBI_tax_id\trelative_abundance\n')
cl2ab, _, tot_nreads = tree.relative_abundances(
pars['tax_lev']+"__" if pars['tax_lev'] != 'a' else None )
fraction_mapped_reads = tot_nreads/float(n_metagenome_reads) if pars['unknown_estimation'] else 1.0
if fraction_mapped_reads > 1.0: fraction_mapped_reads = 1.0
outpred = [(taxstr, taxid,round(relab*100.0,5)) for (taxstr, taxid), relab in cl2ab.items() if relab > 0.0]
if outpred:
if pars['CAMI_format_output']:
for clade, taxid, relab in sorted( outpred, reverse=True,
key=lambda x:x[2]+(100.0*(8-(x[0].count("|"))))):
if taxid:
rank = ranks2code[clade.split('|')[-1][0]]
leaf_taxid = taxid.split('|')[-1]
taxpathsh = '|'.join([remove_prefix(name) if '_unclassified' not in name else '' for name in clade.split('|')])
outf.write( '\t'.join( [ leaf_taxid, rank, taxid, taxpathsh, str(relab*fraction_mapped_reads) ] ) + '\n' )
else:
if pars['unknown_estimation']:
outf.write( "\t".join( [ "UNKNOWN",
"-1",
str(round((1-fraction_mapped_reads)*100,5))]) + "\n" )
for clade, taxid, relab in sorted( outpred, reverse=True,
key=lambda x:x[2]+(100.0*(8-(x[0].count("|"))))):
if not pars['legacy_output']:
outf.write( "\t".join( [clade,
taxid,
str(relab*fraction_mapped_reads)] ) + "\n" )
else:
outf.write( "\t".join( [clade,
str(relab*fraction_mapped_reads)] ) + "\n" )
else:
if not pars['legacy_output']:
outf.write( "UNKNOWN\t-1\t100.0\n" )
else:
outf.write( "UNKNOWN\t100.0\n" )
maybe_generate_biom_file(tree, pars, outpred)
elif pars['t'] == 'rel_ab_w_read_stats':
cl2ab, rr, tot_nreads = tree.relative_abundances(
pars['tax_lev']+"__" if pars['tax_lev'] != 'a' else None )
fraction_mapped_reads = tot_nreads/float(n_metagenome_reads) if not pars['unknown_estimation'] else 1
if fraction_mapped_reads > 1.0: fraction_mapped_reads = 1.0
unmapped_reads = max(n_metagenome_reads - tot_nreads, 0)
outpred = [(taxstr, taxid,round(relab*100.0*fraction_mapped_reads,5)) for (taxstr, taxid),relab in cl2ab.items() if relab > 0.0]
if outpred:
outf.write( "#estimated_reads_mapped_to_known_clades:{}\n".format(tot_nreads) )
outf.write( "\t".join( [ "#clade_name",
"clade_taxid",
"relative_abundance",
"coverage",
"estimated_number_of_reads_from_the_clade" ]) +"\n" )
if not pars['unknown_estimation']:
outf.write( "\t".join( [ "UNKNOWN",
"-1",
str(round((1-fraction_mapped_reads)*100,5)),
"-",
str(unmapped_reads) ]) + "\n" )
for taxstr, taxid, relab in sorted( outpred, reverse=True,
key=lambda x:x[2]+(100.0*(8-(x[0].count("|"))))):
outf.write( "\t".join( [ taxstr,
taxid,
str(relab),
str(round(rr[(taxstr, taxid)][0],5)) if (taxstr, taxid) in rr else '-', #coverage
str( int( round( rr[(taxstr, taxid)][1], 0) ) if (taxstr, taxid) in rr else '-') #estimated_number_of_reads_from_the_clade
] ) + "\n" )
else:
if not pars['legacy_output']:
outf.write( "unclassified\t-1\t100.0\n" )
else:
outf.write( "unclassified\t100.0\n" )
maybe_generate_biom_file(tree, pars, outpred)
elif pars['t'] == 'clade_profiles':
cl2pr = tree.clade_profiles( pars['tax_lev']+"__" if pars['tax_lev'] != 'a' else None )
for c,p in cl2pr.items():
mn,n = zip(*p)
outf.write( "\t".join( [""]+[str(s) for s in mn] ) + "\n" )
outf.write( "\t".join( [c]+[str(s) for s in n] ) + "\n" )
elif pars['t'] == 'marker_ab_table':
cl2pr = tree.clade_profiles( pars['tax_lev']+"__" if pars['tax_lev'] != 'a' else None )
for v in cl2pr.values():
outf.write( "\n".join(["\t".join([str(a),str(b/float(pars['nreads'])) if pars['nreads'] else str(b)])
for a,b in v if b > 0.0]) + "\n" )
elif pars['t'] == 'marker_pres_table':
cl2pr = tree.clade_profiles( pars['tax_lev']+"__" if pars['tax_lev'] != 'a' else None )
for v in cl2pr.values():
strout = ["\t".join([str(a),"1"]) for a,b in v if b > pars['pres_th']]
if strout:
outf.write( "\n".join(strout) + "\n" )
elif pars['t'] == 'marker_counts':
outf.write( "\n".join( ["\t".join([m,str(c)]) for m,c in tree.markers2counts().items() ]) +"\n" )
elif pars['t'] == 'clade_specific_strain_tracker':
cl2pr = tree.clade_profiles( None, get_all = True )
cl2ab, _ = tree.relative_abundances( None )
strout = []
for clade,v in cl2pr.items():
if clade.endswith(pars['clade']) and cl2ab[clade]*100.0 < pars['min_ab']:
strout = []
break
if pars['clade'] in clade:
strout += ["\t".join([str(a),str(int(b > pars['pres_th']))]) for a,b in v]
if strout:
strout = sorted(strout,key=lambda x:x[0])
outf.write( "\n".join(strout) + "\n" )
else:
sys.stderr.write("Clade "+pars['clade']+" not present at an abundance >"+str(round(pars['min_ab'],2))+"%, "
"so no clade specific markers are reported\n")
if __name__ == '__main__':
t0 = time.time()
metaphlan2()
sys.stderr.write('Elapsed time to run MetaPhlAn2: {} s\n'.format( (time.time()-t0) ) )
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