metaphlan2(1)
NAME
metaphlan2 - METAgenomic PHyLogenetic ANalysis for metagenomic taxonomic profiling
SYNOPSIS
metaphlan2
--input_type {fastq,fasta,multifasta,multifastq,bowtie2out,sam}
[--mpa_pkl MPA_PKL] [--bowtie2db METAPHLAN_BOWTIE2_DB] [--bt2_ps
BowTie2 presets] [--bowtie2_exe BOWTIE2_EXE] [--bowtie2out FILE_NAME]
[--no_map] [--tmp_dir] [--tax_lev TAXONOMIC_LEVEL] [--min_cu_len]
[--min_alignment_len] [--ignore_viruses] [--ignore_eukaryotes]
[--ignore_bacteria] [--ignore_archaea] [--stat_q] [--ignore_markers
IGNORE_MARKERS] [--avoid_disqm] [--stat] [-t ANALYSIS TYPE] [--nreads
NUMBER_OF_READS] [--pres_th PRESENCE_THRESHOLD] [--clade] [--min_ab]
[-h] [-o output file] [--sample_id_key name] [--sample_id value] [-s
sam_output_file] [--biom biom_output] [--mdelim mdelim] [--nproc N]
[-v] [INPUT_FILE] [OUTPUT_FILE]
DESCRIPTION
MetaPhlAn 2 clade-abundance estimation
The basic usage of MetaPhlAn 2 consists in the identification of the
clades (from phyla to species and strains in particular cases) present
in the metagenome obtained from a microbiome sample and their relative
abundance. This correspond to the default analysis type
(--analysis_type rel_ab).
* Profiling a metagenome from raw reads:
metaphlan2 metagenome.fastq --input_type fastq
* You can take advantage of multiple CPUs and save the
intermediate BowTie2 output for re-running
MetaPhlAn extremely quickly:
metaphlan2 metagenome.fastq --bowtie2out metagenome.bowtie2.bz2
--nproc 5 --input_type fastq
* If you already mapped your metagenome against the marker DB
(using a previous MetaPhlAn run), you can obtain the results in
few seconds by using the previously saved --bowtie2out file and
specifying the input (--input_type bowtie2out):
metaphlan2 metagenome.bowtie2.bz2 --nproc 5 --input_type
bowtie2out
* You can also provide an externally BowTie2-mapped SAM if you
specify this format with --input_type. Two steps: first apply
BowTie2 and then feed MetaPhlAn2 with the obtained sam:
bowtie2 --sam-no-hd --sam-no-sq --no-unal --very-sensitive -S
metagenome.sam -x /usr/share/metaphlan2/db_v20/mpa_v20_m200 -U
metagenome.fastq metaphlan2 metagenome.sam --input_type sam >
profiled_metagenome.txt
* Multiple alternative ways to pass the input are also available:
cat metagenome.fastq | metaphlan2 --input_type fastq
tar xjf metagenome.tar.bz2 --to-stdout | metaphlan2 --input_type
fastq
metaphlan2 --input_type fastq < metagenome.fastq
metaphlan2 --input_type fastq <(bzcat metagenome.fastq.bz2)
metaphlan2 --input_type fastq <(zcat metagenome_1.fastq.gz
metagenome_2.fastq.gz)
* We can also natively handle paired-end metagenomes, and, more
generally, metagenomes stored in multiple files (but you need to
specify the --bowtie2out parameter):
metaphlan2 metagenome_1.fastq,metagenome_2.fastq --bowtie2out
metagenome.bowtie2.bz2 --nproc 5 --input_type fastq
MetaPhlAn 2 strain tracking
MetaPhlAn 2 introduces the capability of charachterizing organisms at
the strain level using non aggregated marker information. Such
capability comes with several slightly different flavours and are a way
to perform strain tracking and comparison across multiple samples.
Usually, MetaPhlAn 2 is first ran with the default --analysis_type to
profile the species present in the community, and then a strain-level
profiling can be performed to zoom-in into specific species of
interest. This operation can be performed quickly as it exploits the
--bowtie2out intermediate file saved during the execution of the
default analysis type.
* The following command will output the abundance of each marker
with a RPK (reads per kil-base) higher 0.0. (we are assuming
that metagenome_outfmt.bz2 has been generated before as shown
above).
metaphlan2 -t marker_ab_table metagenome_outfmt.bz2 --input_type
bowtie2out > marker_abundance_table.txt
The obtained RPK can be optionally normalized by the total
number of reads in the metagenome to guarantee fair comparisons
of abundances across samples. The number of reads in the
metagenome needs to be passed with the '--nreads' argument
* The list of markers present in the sample can be obtained with
'-t marker_pres_table'
metaphlan2 -t marker_pres_table metagenome_outfmt.bz2
--input_type bowtie2out > marker_abundance_table.txt
The --pres_th argument (default 1.0) set the minimum RPK value
to consider a marker present
* The list '-t clade_profiles' analysis type reports the same
information of '-t marker_ab_table' but the markers are reported
on a clade-by-clade basis.
metaphlan2 -t clade_profiles metagenome_outfmt.bz2 --input_type
bowtie2out > marker_abundance_table.txt
* Finally, to obtain all markers present for a specific clade and
all its subclades, the '-t clade_specific_strain_tracker' should
be used. For example, the following command is reporting the
presence/absence of the markers for the B. fragulis species and
its strains the optional argument --min_ab specifies the minimum
clade abundance for reporting the markers
$ metaphlan2 -t clade_specific_strain_tracker --clade
s__Bacteroides_fragilis metagenome_outfmt.bz2 --input_type
bowtie2out > marker_abundance_table.txt
OPTIONS
positional arguments
INPUT_FILE
the input file can be:
* a fastq file containing metagenomic reads
OR
* a BowTie2 produced SAM file.
OR
* an intermediary mapping file of the metagenome generated by a
previous MetaPhlAn run
If the input file is missing, the script assumes that the input
is provided using the standard input, or named pipes.
IMPORTANT: the type of input needs to be specified with
--input_type
OUTPUT_FILE
the tab-separated output file of the predicted taxon relative
abundances [stdout if not present]
Required arguments
--input_type {fastq,fasta,multifasta,multifastq,bowtie2out,sam}
set whether the input is the multifasta file of metagenomic
reads or the SAM file of the mapping of the reads against the
MetaPhlAn db. [default 'automatic', i.e. the script will try to
guess the input format]
Mapping arguments:
--mpa_pkl MPA_PKL
the metadata pickled MetaPhlAn file
--bowtie2db METAPHLAN_BOWTIE2_DB
The BowTie2 database file of the MetaPhlAn database. Used if
--input_type is fastq, fasta, multifasta, or multifastq
--bt2_ps BowTie2 presets
presets options for BowTie2 (applied only when a multifasta file
is provided) The choices enabled in MetaPhlAn are:
* sensitive
* very-sensitive
* sensitive-local
* very-sensitive-local
[default very-sensitive]
--bowtie2_exe BOWTIE2_EXE
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
--bowtie2out FILE_NAME
The file for saving the output of BowTie2
--no_map
Avoid storing the --bowtie2out map file
--tmp_dir
the folder used to store temporary files [default is the OS
dependent tmp dir]
Post-mapping arguments
--tax_lev TAXONOMIC_LEVEL
The taxonomic level for the relative abundance output:
'a' : all taxonomic levels
'k' : kingdoms
'p' : phyla only
'c' : classes only
'o' : orders only
'f' : families only
'g' : genera only
's' : species only
[default 'a']
--min_cu_len
minimum total nucleotide length for the markers in a clade for
estimating the abundance without considering sub-clade
abundances [default 2000]
--min_alignment_len
The sam records for aligned reads with the longest subalignment
length smaller than this threshold will be discarded. [default
None]
--ignore_viruses
Do not profile viral organisms
--ignore_eukaryotes
Do not profile eukaryotic organisms
--ignore_bacteria
Do not profile bacterial organisms
--ignore_archaea
Do not profile archeal organisms
--stat_q
Quantile value for the robust average [default 0.1]
--ignore_markers IGNORE_MARKERS
File containing a list of markers to ignore.
--avoid_disqm
Deactivate the procedure of disambiguating the quasi-markers
based on the marker abundance pattern found in the sample. It is
generally recommended too keep the disambiguation procedure in
order to minimize false positives
--stat EXPERIMENTAL! Statistical approach for converting marker
abundances into clade abundances
'avg_g' : clade global (i.e. normalizing all markers together)
average
'avg_l' : average of length-normalized marker counts
'tavg_g' : truncated clade global average at --stat_q quantile
'tavg_l' : trunated average of length-normalized marker counts
(at --stat_q)
'wavg_g' : winsorized clade global average (at --stat_q)
'wavg_l' : winsorized average of length-normalized marker counts
(at --stat_q)
'med' : median of length-normalized marker counts
[default tavg_g]
Additional analysis types and arguments
-t ANALYSIS TYPE
Type of analysis to perform:
* rel_ab: profiling a metagenomes in terms of relative abundances
* rel_ab_w_read_stats: profiling a metagenomes in terms of
relative abundances and estimate the number of reads coming from
each clade.
* reads_map: mapping from reads to clades (only reads hitting a
marker)
* clade_profiles: normalized marker counts for clades with at
least a non-null marker
* marker_ab_table: normalized marker counts (only when > 0.0 and
normalized by metagenome size if --nreads is specified)
* marker_counts: non-normalized marker counts [use with extreme
caution]
* marker_pres_table: list of markers present in the sample
(threshold at 1.0 if not differently specified with --pres_th
[default 'rel_ab']
--nreads NUMBER_OF_READS
The total number of reads in the original metagenome. It is used
only when -t marker_table is specified for normalizing the
length-normalized counts with the metagenome size as well. No
normalization applied if --nreads is not specified
--pres_th PRESENCE_THRESHOLD
Threshold for calling a marker present by the -t
marker_pres_table option
--clade
The clade for clade_specific_strain_tracker analysis
--min_ab
The minimum percentage abundace for the clade in the
clade_specific_strain_tracker analysis
-h, --help
show this help message and exit
Output arguments
-o output file, --output_file output file
The output file (if not specified as positional argument)
--sample_id_key name
Specify the sample ID key for this analysis. Defaults to
'#SampleID'.
--sample_id value
Specify the sample ID for this analysis. Defaults to
'Metaphlan2_Analysis'.
-s sam_output_file, --samout sam_output_file
The sam output file
--biom biom_output, --biom_output_file biom_output
If requesting biom file output: The name of the output file in
biom format
--mdelim mdelim, --metadata_delimiter_char mdelim
Delimiter for bug metadata: - defaults to pipe. e.g. the pipe in
k__Bacteria|p__Proteobacteria
Other arguments
--nproc N
The number of CPUs to use for parallelizing the mapping [default
1, i.e. no parallelism]
-v, --version
Prints the current MetaPhlAn version and exit
AUTHOR
The code of MetaPhlAn was rwitten by Nicola Segata (nicola.segata@unitn.it), Duy Tin Truong (duytin.truong@unitn.it). This manpage was written by Andreas Tille for the Debian distribution and can be used for any other usage of the program.
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