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#!/usr/bin/env python3
"""
Create tab-delimited database file to store sequence alignment information
"""
# Info
__author__ = 'Namita Gupta, Jason Anthony Vander Heiden'
from changeo import __version__, __date__
# Imports
import os
import re
import csv
from argparse import ArgumentParser
from collections import OrderedDict
from textwrap import dedent
from time import time
from Bio import SeqIO
# Presto and changeo imports
from presto.Annotation import parseAnnotation
from presto.IO import countSeqFile, printLog, printMessage, printProgress, printError, printWarning, readSeqFile
from changeo.Defaults import default_format, default_out_args, default_imgt_id_len
from changeo.Commandline import CommonHelpFormatter, checkArgs, getCommonArgParser, parseCommonArgs
from changeo.Alignment import RegionDefinition, gapV
from changeo.Gene import buildGermline
from changeo.IO import countDbFile, extractIMGT, readGermlines, getFormatOperators, getOutputHandle, \
AIRRWriter, ChangeoWriter, IgBLASTReader, IgBLASTReaderAA, IMGTReader, IHMMuneReader, \
checkFields
from changeo.Receptor import ChangeoSchema, AIRRSchema
# 10X Receptor attributes
cellranger_base = ['cell', 'c_call', 'conscount', 'umicount']
cellranger_extended = ['cell', 'c_call', 'conscount', 'umicount',
'v_call_10x', 'd_call_10x', 'j_call_10x',
'junction_10x', 'junction_10x_aa']
def readCellRanger(cellranger_file, fields=cellranger_base):
"""
Load a Cell Ranger annotation table
Arguments:
cellranger_file (str): path to the annotation file.
fields (list): list of fields to keep.
Returns:
dict: dict of dicts with contig_id as the primary key.
"""
# Mapping of 10X annotations to Receptor attributes
cellranger_map = {'cell': 'barcode',
'c_call': 'c_gene',
'locus': 'chain',
'conscount': 'reads',
'umicount': 'umis',
'v_call_10x': 'v_gene',
'd_call_10x': 'd_gene',
'j_call_10x': 'j_gene',
'junction_10x': 'cdr3_nt',
'junction_10x_aa': 'cdr3'}
# Function to parse individual fields
def _parse(x):
return '' if x == 'None' else x
# Generate annotation dictionary
ann_dict = {}
with open(cellranger_file) as csv_file:
# Detect delimiters
dialect = csv.Sniffer().sniff(csv_file.readline())
csv_file.seek(0)
# Read in annotation file
csv_reader = csv.DictReader(csv_file, dialect=dialect)
# Generate annotation dictionary
for row in csv_reader:
ann_dict[row['contig_id']] = {f: _parse(row[cellranger_map[f]]) for f in fields}
return ann_dict
def addGermline(receptor, references, amino_acid=False):
"""
Add full length germline to Receptor object
Arguments:
receptor (changeo.Receptor.Receptor): Receptor object to modify.
references (dict): dictionary of IMGT-gapped references sequences.
amino_acid (bool): if True build amino acid germline, otherwise build nucleotide germline
Returns:
changeo.Receptor.Receptor: modified Receptor with the germline sequence added.
"""
if amino_acid:
__, germlines, __ = buildGermline(receptor, references, seq_field='sequence_aa_imgt',
amino_acid=True)
germline_seq = None if germlines is None else germlines['full']
receptor.setField('germline_aa_imgt', germline_seq)
else:
__, germlines, __ = buildGermline(receptor, references, amino_acid=False)
germline_seq = None if germlines is None else germlines['full']
receptor.setField('germline_imgt', germline_seq)
return receptor
def getIDforIMGT(seq_file, imgt_id_len=default_imgt_id_len):
"""
Create a sequence ID translation using IMGT truncation.
Arguments:
seq_file (str): a fasta file of sequences input to IMGT.
Returns:
dict: a dictionary of with the IMGT truncated ID as the key and the full sequence description as the value.
"""
# Create a sequence ID translation using IDs truncate up to space or 49 chars
ids = {}
for rec in readSeqFile(seq_file):
if len(rec.description) <= imgt_id_len:
id_key = rec.description
else: # truncate and replace characters
if imgt_id_len == 49: # 28 September 2021 (version 1.8.4)
id_key = re.sub('\s|\t', '_', rec.description[:imgt_id_len])
else: # older versions
id_key = re.sub('\||\s|!|&|\*|<|>|\?', '_', rec.description[:imgt_id_len])
ids.update({id_key: rec.description})
return ids
def correctIMGTFields(receptor, references):
"""
Add IMGT-numbering to IMGT fields in a Receptor object
Arguments:
receptor (changeo.Receptor.Receptor): Receptor object to modify.
references (dict): dictionary of IMGT-gapped references sequences.
Returns:
changeo.Receptor.Receptor: modified Receptor with IMGT-gapped fields.
"""
# Initialize update object
imgt_dict = {'sequence_imgt': None,
'v_germ_start_imgt': None,
'v_germ_length_imgt': None,
'germline_imgt': None}
# Check for necessary fields
try:
if not all([receptor.sequence_imgt,
receptor.v_germ_start_imgt,
receptor.v_germ_length_imgt,
receptor.v_call]):
raise AttributeError
except AttributeError:
return None
# Gap V region
try:
gapped = gapV(receptor.sequence_imgt,
receptor.v_germ_start_imgt,
receptor.v_germ_length_imgt,
receptor.v_call,
references)
except KeyError as e:
printWarning(e)
return None
# Verify IMGT-gapped sequence and junction concur
# try:
# check = (receptor.junction == gapped['sequence_imgt'][309:(309 + receptor.junction_length)])
# except TypeError:
# check = False
# if not check:
# return None
# Rebuild germline sequence
receptor.setDict(gapped, parse=False)
__, germlines, __ = buildGermline(receptor, references)
# log, germlines, genes = buildGermline(receptor, references)
# print(log)
if germlines is not None:
gapped['germline_imgt'] = germlines['full']
else:
return None
# Update return object
imgt_dict.update(gapped)
return imgt_dict
def getSeqDict(seq_file):
"""
Create a dictionary from a sequence file.
Arguments:
seq_file (str): sequence file.
Returns:
dict: sequence description as keys with Bio.SeqRecords as values.
"""
seq_dict = SeqIO.to_dict(readSeqFile(seq_file), key_function=lambda x: x.description)
return seq_dict
def writeDb(records, fields, aligner_file, total_count, id_dict=None, annotations=None,
amino_acid=False, validate='strict', asis_id=True, regions='default',
writer=AIRRWriter, out_file=None, out_args=default_out_args):
"""
Writes parsed records to an output file
Arguments:
records (iter): a iterator of Receptor objects containing alignment data.
fields (list): a list of ordered field names to write.
aligner_file (str): input file name.
total_count (int): number of records (for progress bar).
id_dict (dict): a dictionary of the truncated sequence ID mapped to the full sequence ID.
annotations (dict): additional annotation dictionary.
amino_acid (bool): if True do verification on amino acid fields.
validate (str): validation criteria for passing records; one of 'strict', 'gentle', or 'partial'.
asis_id (bool): if ID is to be parsed for pRESTO output with default delimiters.
regions (str): name of the IMGT FWR/CDR region definitions to use.
writer (changeo.IO.TSVWriter): writer class.
out_file (str): output file name. Automatically generated from the input file if None.
out_args (dict): common output argument dictionary from parseCommonArgs.
Returns:
None
"""
# Wrapper for opening handles and writers
def _open(x, f, writer=writer, out_file=out_file):
if out_file is not None and x == 'pass':
handle = open(out_file, 'w')
else:
handle = getOutputHandle(aligner_file,
out_label='db-%s' % x,
out_dir=out_args['out_dir'],
out_name=out_args['out_name'],
out_type=out_args['out_type'])
return handle, writer(handle, fields=f)
# Function to convert fasta header annotations to changeo columns
def _changeo(f, header):
h = [ChangeoSchema.fromReceptor(x) for x in header if x.upper() not in f]
f.extend(h)
return f
def _airr(f, header):
h = [AIRRSchema.fromReceptor(x) for x in header if x.lower() not in f]
f.extend(h)
return f
# Function to verify IMGT-gapped sequence and junction concur
def _imgt_check(rec):
try:
if amino_acid:
rd = RegionDefinition(rec.junction_aa_length, amino_acid=amino_acid, definition=regions)
x, y = rd.positions['junction']
check = (rec.junction_aa == rec.sequence_aa_imgt[x:y])
else:
rd = RegionDefinition(rec.junction_length, amino_acid=amino_acid, definition=regions)
x, y = rd.positions['junction']
check = (rec.junction == rec.sequence_imgt[x:y])
except (TypeError, AttributeError):
check = False
return check
# Default function to check for valid records
def _gentle(rec):
if amino_acid:
valid = [rec.v_call and rec.v_call != 'None',
rec.j_call and rec.j_call != 'None',
rec.functional is not None,
rec.sequence_aa_imgt,
rec.junction_aa]
else:
valid = [rec.v_call and rec.v_call != 'None',
rec.j_call and rec.j_call != 'None',
rec.functional is not None,
rec.sequence_imgt,
rec.junction]
return all(valid)
# Function to check for valid records strictly
def _strict(rec):
if amino_acid:
valid = [rec.v_call and rec.v_call != 'None',
rec.j_call and rec.j_call != 'None',
rec.functional is not None,
rec.sequence_aa_imgt,
rec.junction_aa,
_imgt_check(rec)]
else:
valid = [rec.v_call and rec.v_call != 'None',
rec.j_call and rec.j_call != 'None',
rec.functional is not None,
rec.sequence_imgt,
rec.junction,
_imgt_check(rec)]
return all(valid)
# Function to check for valid records loosely
def _partial(rec):
valid = [rec.v_call and rec.v_call != 'None',
rec.d_call and rec.d_call != 'None',
rec.j_call and rec.j_call != 'None']
return any(valid)
# Set writer class and annotation conversion function
if writer == ChangeoWriter:
_annotate = _changeo
elif writer == AIRRWriter:
_annotate = _airr
else:
printError('Invalid output writer.')
# Set pass criteria
validate_map = {'strict': _strict, 'gentle': _gentle, 'partial': _partial}
_pass = validate_map.get(validate, _gentle)
# Define log handle
if out_args['log_file'] is None:
log_handle = None
else:
log_handle = open(out_args['log_file'], 'w')
# Initialize handles, writers and counters
pass_handle, pass_writer = None, None
fail_handle, fail_writer = None, None
pass_count, fail_count = 0, 0
start_time = time()
# Validate and write output
printProgress(0, total_count, 0.05, start_time=start_time)
for i, record in enumerate(records, start=1):
# Replace sequence description with full string, if required
if id_dict is not None and record.sequence_id in id_dict:
record.sequence_id = id_dict[record.sequence_id]
# Parse sequence description into new columns
if not asis_id:
try:
ann_raw = parseAnnotation(record.sequence_id)
record.sequence_id = ann_raw.pop('ID')
# Convert to Receptor fields
ann_parsed = OrderedDict()
for k, v in ann_raw.items():
ann_parsed[ChangeoSchema.toReceptor(k)] = v
# Add annotations to Receptor and update field list
record.setDict(ann_parsed, parse=True)
if i == 1: fields = _annotate(fields, ann_parsed.keys())
except IndexError:
# Could not parse pRESTO-style annotations so fall back to no parse
asis_id = True
printWarning('Sequence annotation format not recognized. Sequence headers will not be parsed.')
# Add supplemental annotation fields
# if _append_table is not None:
# record.setDict(_append_table(record.sequence_id), parse=True)
if annotations is not None:
record.setDict(annotations[record.sequence_id], parse=True)
if i == 1: fields = _annotate(fields, annotations[record.sequence_id].keys())
# Count pass or fail and write to appropriate file
if _pass(record):
pass_count += 1
# Write row to pass file
try:
pass_writer.writeReceptor(record)
except AttributeError:
# Open pass file and writer
pass_handle, pass_writer = _open('pass', fields)
pass_writer.writeReceptor(record)
else:
fail_count += 1
# Write row to fail file if specified
if out_args['failed']:
try:
fail_writer.writeReceptor(record)
except AttributeError:
# Open fail file and writer
fail_handle, fail_writer = _open('fail', fields)
fail_writer.writeReceptor(record)
# Write log
if log_handle is not None:
log = OrderedDict([('ID', record.sequence_id),
('V_CALL', record.v_call),
('D_CALL', record.d_call),
('J_CALL', record.j_call),
('PRODUCTIVE', record.functional)])
if not _imgt_check(record) and not amino_acid:
log['ERROR'] = 'Junction does not match the sequence starting at position 310 in the IMGT numbered V(D)J sequence.'
printLog(log, log_handle)
# Print progress
printProgress(i, total_count, 0.05, start_time=start_time)
# Print console log
log = OrderedDict()
log['OUTPUT'] = os.path.basename(pass_handle.name) if pass_handle is not None else None
log['PASS'] = pass_count
log['FAIL'] = fail_count
log['END'] = 'MakeDb'
printLog(log)
# Close file handles
output = {'pass': None, 'fail': None}
if pass_handle is not None:
output['pass'] = pass_handle.name
pass_handle.close()
if fail_handle is not None:
output['fail'] = fail_handle.name
fail_handle.close()
return output
def parseIMGT(aligner_file, seq_file=None, repo=None, cellranger_file=None, validate='strict', asis_id=True,
extended=False, format=default_format, out_file=None, out_args=default_out_args,
imgt_id_len=default_imgt_id_len):
"""
Main for IMGT aligned sample sequences.
Arguments:
aligner_file (str): zipped file or unzipped folder output by IMGT.
seq_file (str): FASTA file input to IMGT (from which to get seqID).
repo (str): folder with germline repertoire files.
validate (str): validation criteria for passing records; one of 'strict', 'gentle', or 'partial'.
asis_id (bool): if ID is to be parsed for pRESTO output with default delimiters.
extended (bool): if True add alignment score, FWR, CDR and junction fields to output file.
format (str): output format. one of 'changeo' or 'airr'.
out_file (str): output file name. Automatically generated from the input file if None.
out_args (dict): common output argument dictionary from parseCommonArgs.
imgt_id_len (int): maximum character length of sequence identifiers reported by IMGT/HighV-QUEST.
Returns:
dict: names of the 'pass' and 'fail' output files.
"""
# Print parameter info
log = OrderedDict()
log['START'] = 'MakeDb'
log['COMMAND'] = 'imgt'
log['ALIGNER_FILE'] = aligner_file
log['SEQ_FILE'] = os.path.basename(seq_file) if seq_file else ''
log['ASIS_ID'] = asis_id
log['VALIDATE'] = validate
log['EXTENDED'] = extended
printLog(log)
start_time = time()
printMessage('Loading files', start_time=start_time, width=20)
# Extract IMGT files
temp_dir, imgt_files = extractIMGT(aligner_file)
# Count records in IMGT files
total_count = countDbFile(imgt_files['summary'])
# Get (parsed) IDs from fasta file submitted to IMGT
id_dict = getIDforIMGT(seq_file, imgt_id_len) if seq_file else {}
# Load supplementary annotation table
if cellranger_file is not None:
f = cellranger_extended if extended else cellranger_base
annotations = readCellRanger(cellranger_file, fields=f)
else:
annotations = None
printMessage('Done', start_time=start_time, end=True, width=20)
# Define format operators
try:
__, writer, schema = getFormatOperators(format)
except ValueError:
printError('Invalid format %s.' % format)
out_args['out_type'] = schema.out_type
# Define output fields
fields = list(schema.required)
if extended:
custom = IMGTReader.customFields(scores=True, regions=True, junction=True, schema=schema)
fields.extend(custom)
# Parse IMGT output and write db
with open(imgt_files['summary'], 'r') as summary_handle, \
open(imgt_files['gapped'], 'r') as gapped_handle, \
open(imgt_files['ntseq'], 'r') as ntseq_handle, \
open(imgt_files['junction'], 'r') as junction_handle:
# Open parser
parse_iter = IMGTReader(summary_handle, gapped_handle, ntseq_handle, junction_handle)
# Add germline sequence
if repo is None:
germ_iter = parse_iter
else:
references = readGermlines(repo)
# Check for IMGT-gaps in germlines
if all('...' not in x for x in references.values()):
printWarning('Germline reference sequences do not appear to contain IMGT-numbering spacers. Results may be incorrect.')
germ_iter = (addGermline(x, references) for x in parse_iter)
# Write db
output = writeDb(germ_iter, fields=fields, aligner_file=aligner_file, total_count=total_count,
annotations=annotations, id_dict=id_dict, asis_id=asis_id, validate=validate,
writer=writer, out_file=out_file, out_args=out_args)
# Cleanup temp directory
temp_dir.cleanup()
return output
def parseIgBLAST(aligner_file, seq_file, repo, amino_acid=False, cellranger_file=None, validate='strict',
asis_id=True, asis_calls=False, extended=False, regions='default', infer_junction=False,
format='changeo', out_file=None, out_args=default_out_args):
"""
Main for IgBLAST aligned sample sequences.
Arguments:
aligner_file (str): IgBLAST output file to process.
seq_file (str): fasta file input to IgBlast (from which to get sequence).
repo (str): folder with germline repertoire files.
amino_acid (bool): if True then the IgBLAST output files are results from igblastp. igblastn is assumed if False.
validate (str): validation criteria for passing records; one of 'strict', 'gentle', or 'partial'.
asis_id (bool): if ID is to be parsed for pRESTO output with default delimiters.
asis_calls (bool): if True do not parse gene calls for allele names.
extended (bool): if True add alignment scores, FWR regions, and CDR regions to the output.
regions (str): name of the IMGT FWR/CDR region definitions to use.
infer_junction (bool): if True, infer the junction sequence, if not reported by IgBLAST.
format (str): output format. one of 'changeo' or 'airr'.
out_file (str): output file name. Automatically generated from the input file if None.
out_args (dict): common output argument dictionary from parseCommonArgs.
Returns:
dict: names of the 'pass' and 'fail' output files.
"""
# Print parameter info
log = OrderedDict()
log['START'] = 'MakeDB'
log['COMMAND'] = 'igblast-aa' if amino_acid else 'igblast'
log['ALIGNER_FILE'] = os.path.basename(aligner_file)
log['SEQ_FILE'] = os.path.basename(seq_file)
log['ASIS_ID'] = asis_id
log['ASIS_CALLS'] = asis_calls
log['VALIDATE'] = validate
log['EXTENDED'] = extended
log['INFER_JUNCTION'] = infer_junction
printLog(log)
# Set amino acid conditions
if amino_acid:
format = '%s-aa' % format
parser = IgBLASTReaderAA
else:
parser = IgBLASTReader
# Start
start_time = time()
printMessage('Loading files', start_time=start_time, width=20)
# Count records in sequence file
total_count = countSeqFile(seq_file)
# Get input sequence dictionary
seq_dict = getSeqDict(seq_file)
# Create germline repo dictionary
references = readGermlines(repo, asis=asis_calls)
# Load supplementary annotation table
if cellranger_file is not None:
f = cellranger_extended if extended else cellranger_base
annotations = readCellRanger(cellranger_file, fields=f)
else:
annotations = None
printMessage('Done', start_time=start_time, end=True, width=20)
# Check for IMGT-gaps in germlines
if all('...' not in x for x in references.values()):
printWarning('Germline reference sequences do not appear to contain IMGT-numbering spacers. Results may be incorrect.')
# Define format operators
try:
__, writer, schema = getFormatOperators(format)
except ValueError:
printError('Invalid format %s.' % format)
out_args['out_type'] = schema.out_type
# Define output fields
fields = list(schema.required)
if extended:
custom = parser.customFields(schema=schema)
fields.extend(custom)
# Parse and write output
with open(aligner_file, 'r') as f:
parse_iter = parser(f, seq_dict, references, regions=regions, asis_calls=asis_calls, infer_junction=infer_junction)
germ_iter = (addGermline(x, references, amino_acid=amino_acid) for x in parse_iter)
output = writeDb(germ_iter, fields=fields, aligner_file=aligner_file, total_count=total_count,
annotations=annotations, amino_acid=amino_acid, validate=validate, asis_id=asis_id,
regions=regions, writer=writer, out_file=out_file, out_args=out_args)
return output
def parseIHMM(aligner_file, seq_file, repo, cellranger_file=None, validate='strict', asis_id=True,
extended=False, format=default_format, out_file=None, out_args=default_out_args):
"""
Main for iHMMuneAlign aligned sample sequences.
Arguments:
aligner_file (str): iHMMune-Align output file to process.
seq_file (str): fasta file input to iHMMuneAlign (from which to get sequence).
repo (str): folder with germline repertoire files.
validate (str): validation criteria for passing records; one of 'strict', 'gentle', or 'partial'.
asis_id (bool): if ID is to be parsed for pRESTO output with default delimiters.
extended (bool): if True parse alignment scores, FWR and CDR region fields.
format (str): output format. One of 'changeo' or 'airr'.
out_file (str): output file name. Automatically generated from the input file if None.
out_args (dict): common output argument dictionary from parseCommonArgs.
Returns:
dict: names of the 'pass' and 'fail' output files.
"""
# Print parameter info
log = OrderedDict()
log['START'] = 'MakeDB'
log['COMMAND'] = 'ihmm'
log['ALIGNER_FILE'] = os.path.basename(aligner_file)
log['SEQ_FILE'] = os.path.basename(seq_file)
log['ASIS_ID'] = asis_id
log['VALIDATE'] = validate
log['EXTENDED'] = extended
printLog(log)
start_time = time()
printMessage('Loading files', start_time=start_time, width=20)
# Count records in sequence file
total_count = countSeqFile(seq_file)
# Get input sequence dictionary
seq_dict = getSeqDict(seq_file)
# Create germline repo dictionary
references = readGermlines(repo)
# Load supplementary annotation table
if cellranger_file is not None:
f = cellranger_extended if extended else cellranger_base
annotations = readCellRanger(cellranger_file, fields=f)
else:
annotations = None
printMessage('Done', start_time=start_time, end=True, width=20)
# Check for IMGT-gaps in germlines
if all('...' not in x for x in references.values()):
printWarning('Germline reference sequences do not appear to contain IMGT-numbering spacers. Results may be incorrect.')
# Define format operators
try:
__, writer, schema = getFormatOperators(format)
except ValueError:
printError('Invalid format %s.' % format)
out_args['out_type'] = schema.out_type
# Define output fields
fields = list(schema.required)
if extended:
custom = IHMMuneReader.customFields(scores=True, regions=True, schema=schema)
fields.extend(custom)
# Parse and write output
with open(aligner_file, 'r') as f:
parse_iter = IHMMuneReader(f, seq_dict, references)
germ_iter = (addGermline(x, references) for x in parse_iter)
output = writeDb(germ_iter, fields=fields, aligner_file=aligner_file, total_count=total_count,
annotations=annotations, asis_id=asis_id, validate=validate,
writer=writer, out_file=out_file, out_args=out_args)
return output
def numberAIRR(aligner_file, repo=None, format=default_format,
out_file=None, out_args=default_out_args):
"""
Inserts IMGT numbering into V fields
Arguments:
aligner_file (str): AIRR Rearrangement file from the alignment tool.
repo (str): folder with germline repertoire files. If None, do not updated alignment columns with IMGT gaps.
format (str): output format.
out_file (str): output file name. Automatically generated from the input file if None.
out_args (dict): common output argument dictionary from parseCommonArgs.
Returns:
str: output file name.
"""
log = OrderedDict()
log['START'] = 'MakeDb'
log['COMMAND'] = 'number'
log['ALIGNER_FILE'] = os.path.basename(aligner_file)
printLog(log)
# Define format operators
try:
reader, writer, schema = getFormatOperators(format)
except ValueError:
printError('Invalid format %s.' % format)
# Open input
db_handle = open(aligner_file, 'rt')
db_iter = reader(db_handle)
# Define log handle
if out_args['log_file'] is None:
log_handle = None
else:
log_handle = open(out_args['log_file'], 'w')
# Check for required columns
try:
required = ['sequence_imgt', 'v_germ_start_imgt']
checkFields(required, db_iter.fields, schema=schema)
except LookupError as e:
printError(e)
# Load references
reference_dict = readGermlines(repo)
# Check for IMGT-gaps in germlines
if all('...' not in x for x in reference_dict.values()):
printWarning('Germline reference sequences do not appear to contain IMGT-numbering spacers. Results may be incorrect.')
# Open output writer
if out_file is not None:
pass_handle = open(out_file, 'w')
else:
pass_handle = getOutputHandle(aligner_file, out_label='db-pass', out_dir=out_args['out_dir'],
out_name=out_args['out_name'], out_type=schema.out_type)
pass_writer = writer(pass_handle, fields=db_iter.fields)
if out_args['failed']:
fail_handle = getOutputHandle(aligner_file, out_label='db-fail', out_dir=out_args['out_dir'],
out_name=out_args['out_name'], out_type=schema.out_type)
fail_writer = writer(fail_handle, fields=db_iter.fields)
# Count records
result_count = countDbFile(aligner_file)
# Iterate over records
start_time = time()
rec_count = pass_count = fail_count= 0
for rec in db_iter:
# Print progress for previous iteration
printProgress(rec_count, result_count, 0.05, start_time=start_time)
rec_count += 1
# Update IMGT fields
imgt_dict = correctIMGTFields(rec, reference_dict)
# Write records
if imgt_dict is not None:
pass_count += 1
rec.setDict(imgt_dict, parse=False)
pass_writer.writeReceptor(rec)
else:
fail_count += 1
# Write row to fail file if specified
if out_args['failed']:
fail_writer.writeReceptor(rec)
# Write log
if log_handle is not None:
log = OrderedDict([('ID', rec.sequence_id),
('V_CALL', rec.v_call),
('D_CALL', rec.d_call),
('J_CALL', rec.j_call),
('PRODUCTIVE', rec.functional)])
printLog(log, log_handle)
# Print counts
printProgress(rec_count, result_count, 0.05, start_time=start_time)
log = OrderedDict()
log['OUTPUT'] = os.path.basename(pass_handle.name)
log['RECORDS'] = rec_count
log['PASS'] = pass_count
log['FAIL'] = rec_count - pass_count
log['END'] = 'MakeDb'
printLog(log)
# Close file handles
pass_handle.close()
db_handle.close()
return pass_handle.name
def getArgParser():
"""
Defines the ArgumentParser.
Returns:
argparse.ArgumentParser
"""
fields = dedent(
'''
output files:
db-pass
database of alignment records with functionality information,
V and J calls, and a junction region.
db-fail
database with records that fail due to no productivity information,
no gene V assignment, no J assignment, or no junction region.
universal output fields:
sequence_id, sequence, sequence_alignment, germline_alignment,
rev_comp, productive, stop_codon, vj_in_frame, locus,
v_call, d_call, j_call, c_call, junction, junction_length, junction_aa,
v_sequence_start, v_sequence_end, v_germline_start, v_germline_end,
d_sequence_start, d_sequence_end, d_germline_start, d_germline_end,
j_sequence_start, j_sequence_end, j_germline_start, j_germline_end,
np1_length, np2_length, fwr1, fwr2, fwr3, fwr4, cdr1, cdr2, cdr3
imgt specific output fields:
n1_length, n2_length, p3v_length, p5d_length, p3d_length, p5j_length,
d_frame, v_score, v_identity, d_score, d_identity, j_score, j_identity
igblast specific output fields:
v_score, v_identity, v_support, v_cigar,
d_score, d_identity, d_support, d_cigar,
j_score, j_identity, j_support, j_cigar
ihmm specific output fields:
vdj_score
10x specific output fields:
cell_id, consensus_count, umi_count,
v_call_10x, d_call_10x, j_call_10x,
junction_10x, junction_10x_aa
''')
# Define ArgumentParser
parser = ArgumentParser(description=__doc__, epilog=fields,
formatter_class=CommonHelpFormatter, add_help=False)
group_help = parser.add_argument_group('help')
group_help.add_argument('--version', action='version',
version='%(prog)s:' + ' %s %s' %(__version__, __date__))
group_help.add_argument('-h', '--help', action='help', help='show this help message and exit')
subparsers = parser.add_subparsers(title='subcommands', dest='command',
help='Aligner used', metavar='')
# TODO: This is a temporary fix for Python issue 9253
subparsers.required = True
# Parent parser
parser_parent = getCommonArgParser(db_in=False)
# igblastn output parser
parser_igblast = subparsers.add_parser('igblast', parents=[parser_parent],
formatter_class=CommonHelpFormatter, add_help=False,
help='Process igblastn output.',
description='Process igblastn output.')
group_igblast = parser_igblast.add_argument_group('aligner parsing arguments')
group_igblast.add_argument('-i', nargs='+', action='store', dest='aligner_files', required=True,
help='''IgBLAST output files in format 7 with query sequence
(igblastn argument \'-outfmt "7 std qseq sseq btop"\').''')
group_igblast.add_argument('-r', nargs='+', action='store', dest='repo', required=True,
help='''List of folders and/or fasta files containing
the same germline set used in the IgBLAST alignment. These
reference sequences must contain IMGT-numbering spacers (gaps)
in the V segment.''')
group_igblast.add_argument('-s', action='store', nargs='+', dest='seq_files',
required=True,
help='''List of input FASTA files (with .fasta, .fna or .fa
extension), containing sequences.''')
group_igblast.add_argument('--10x', action='store', nargs='+', dest='cellranger_files',
help='''Table file containing 10X annotations (with .csv or .tsv
extension).''')
group_igblast.add_argument('--asis-id', action='store_true', dest='asis_id',
help='''Specify to prevent input sequence headers from being parsed
to add new columns to database. Parsing of sequence headers requires
headers to be in the pRESTO annotation format, so this should be specified
when sequence headers are incompatible with the pRESTO annotation scheme.
Note, unrecognized header formats will default to this behavior.''')
group_igblast.add_argument('--asis-calls', action='store_true', dest='asis_calls',
help='''Specify to prevent gene calls from being parsed into standard allele names
in both the IgBLAST output and reference database. Note, this requires
the sequence identifiers in the reference sequence set and the IgBLAST
database to be exact string matches.''')
group_igblast.add_argument('--extended', action='store_true', dest='extended',
help='''Specify to include additional aligner specific fields in the output.
Adds <vdj>_score, <vdj>_identity, <vdj>_support, <vdj>_cigar,
fwr1, fwr2, fwr3, fwr4, cdr1, cdr2 and cdr3.''')
group_igblast.add_argument('--regions', action='store', dest='regions',
choices=('default', 'rhesus-igl'), default='default',
help='''IMGT CDR and FWR boundary definition to use.''')
group_igblast.add_argument('--infer-junction', action='store_true', dest='infer_junction',
help='''Infer the junction sequence. For use with IgBLAST v1.6.0 or older,
prior to the addition of IMGT-CDR3 inference.''')
group_igblast_validate = group_igblast.add_mutually_exclusive_group(required=False)
# group_igblast_validate.add_argument('--strict', action='store_const', const='strict', dest='validate',
# help='''By default, passing records must contain valid values for the
# V gene, J gene, junction region, and productivity call. If specified,
# this argument adds the additional requirement that the junction region must
# start at position 310 in the IMGT-numbered sequence.''')
group_igblast_validate.add_argument('--partial', action='store_const', const='partial', dest='validate',
help='''If specified, include incomplete V(D)J alignments in
the pass file instead of the fail file. An incomplete alignment
is defined as a record that is missing a V gene assignment,
J gene assignment, junction region, or productivity call.''')
parser_igblast.set_defaults(func=parseIgBLAST, amino_acid=False, validate='strict')
# igblastp output parser
parser_igblast_aa = subparsers.add_parser('igblast-aa', parents=[parser_parent],
formatter_class=CommonHelpFormatter, add_help=False,
help='Process igblastp output.',
description='Process igblastp output.')
group_igblast_aa = parser_igblast_aa.add_argument_group('aligner parsing arguments')
group_igblast_aa.add_argument('-i', nargs='+', action='store', dest='aligner_files', required=True,
help='''IgBLAST output files in format 7 with query sequence
(igblastp argument \'-outfmt "7 std qseq sseq btop"\').''')
group_igblast_aa.add_argument('-r', nargs='+', action='store', dest='repo', required=True,
help='''List of folders and/or fasta files containing
the same germline set used in the IgBLAST alignment. These
reference sequences must contain IMGT-numbering spacers (gaps)
in the V segment.''')
group_igblast_aa.add_argument('-s', action='store', nargs='+', dest='seq_files', required=True,
help='''List of input FASTA files (with .fasta, .fna or .fa
extension), containing sequences.''')
group_igblast_aa.add_argument('--10x', action='store', nargs='+', dest='cellranger_files',
help='''Table file containing 10X annotations (with .csv or .tsv extension).''')
group_igblast_aa.add_argument('--asis-id', action='store_true', dest='asis_id',
help='''Specify to prevent input sequence headers from being parsed
to add new columns to database. Parsing of sequence headers requires
headers to be in the pRESTO annotation format, so this should be specified
when sequence headers are incompatible with the pRESTO annotation scheme.
Note, unrecognized header formats will default to this behavior.''')
group_igblast_aa.add_argument('--asis-calls', action='store_true', dest='asis_calls',
help='''Specify to prevent gene calls from being parsed into standard allele names
in both the IgBLAST output and reference database. Note, this requires
the sequence identifiers in the reference sequence set and the IgBLAST
database to be exact string matches.''')
group_igblast_aa.add_argument('--extended', action='store_true', dest='extended',
help='''Specify to include additional aligner specific fields in the output.
Adds v_score, v_identity, v_support, v_cigar, fwr1, fwr2, fwr3, cdr1 and cdr2.''')
group_igblast_aa.add_argument('--regions', action='store', dest='regions',
choices=('default', 'rhesus-igl'), default='default',
help='''IMGT CDR and FWR boundary definition to use.''')
parser_igblast_aa.set_defaults(func=parseIgBLAST, amino_acid=True, validate='strict')
# IMGT aligner
parser_imgt = subparsers.add_parser('imgt', parents=[parser_parent],
formatter_class=CommonHelpFormatter, add_help=False,
help='''Process IMGT/HighV-Quest output
(does not work with V-QUEST).''',
description='''Process IMGT/HighV-Quest output
(does not work with V-QUEST).''')
group_imgt = parser_imgt.add_argument_group('aligner parsing arguments')
group_imgt.add_argument('-i', nargs='+', action='store', dest='aligner_files', required=True,
help='''Either zipped IMGT output files (.zip or .txz) or a
folder containing unzipped IMGT output files (which must
include 1_Summary, 2_IMGT-gapped, 3_Nt-sequences,
and 6_Junction).''')
group_imgt.add_argument('-s', nargs='*', action='store', dest='seq_files', required=False,
help='''List of FASTA files (with .fasta, .fna or .fa
extension) that were submitted to IMGT/HighV-QUEST.
If unspecified, sequence identifiers truncated by IMGT/HighV-QUEST
will not be corrected.''')
group_imgt.add_argument('-r', nargs='+', action='store', dest='repo', required=False,
help='''List of folders and/or fasta files containing
the germline sequence set used by IMGT/HighV-QUEST.
These reference sequences must contain IMGT-numbering spacers (gaps)
in the V segment. If unspecified, the germline sequence reconstruction
will not be included in the output.''')
group_imgt.add_argument('--10x', action='store', nargs='+', dest='cellranger_files',
help='''Table file containing 10X annotations (with .csv or .tsv
extension).''')
group_imgt.add_argument('--extended', action='store_true', dest='extended',
help='''Specify to include additional aligner specific fields in the output.
Adds <vdj>_score, <vdj>_identity>, fwr1, fwr2, fwr3, fwr4,
cdr1, cdr2, cdr3, n1_length, n2_length, p3v_length, p5d_length,
p3d_length, p5j_length and d_frame.''')
group_imgt.add_argument('--asis-id', action='store_true', dest='asis_id',
help='''Specify to prevent input sequence headers from being parsed
to add new columns to database. Parsing of sequence headers requires
headers to be in the pRESTO annotation format, so this should be specified
when sequence headers are incompatible with the pRESTO annotation scheme.
Note, unrecognized header formats will default to this behavior.''')
group_imgt.add_argument('--imgt-id-len', action='store', dest='imgt_id_len', type=int,
default=default_imgt_id_len,
help='''The maximum character length of sequence identifiers reported by IMGT/HighV-QUEST.
Specify 50 if the IMGT files (-i) were generated with an IMGT/HighV-QUEST version older
than 1.8.3 (May 7, 2021).''')
group_imgt_validate = group_imgt.add_mutually_exclusive_group(required=False)
# group_imgt_validate.add_argument('--strict', action='store_const', const='strict', dest='validate',
# help='''By default, passing records must contain valid values for the
# V gene, J gene, junction region, and productivity call. If specified,
# this argument adds the additional requirement that the junction region must
# start at position 310 in the IMGT-numbered sequence.''')
group_imgt_validate.add_argument('--partial', action='store_const', const='partial', dest='validate',
help='''If specified, include incomplete V(D)J alignments in
the pass file instead of the fail file. An incomplete alignment
is defined as a record that is missing a V gene assignment,
J gene assignment, junction region, or productivity call.''')
parser_imgt.set_defaults(func=parseIMGT, validate='strict')
# iHMMuneAlign Aligner
parser_ihmm = subparsers.add_parser('ihmm', parents=[parser_parent],
formatter_class=CommonHelpFormatter, add_help=False,
help='Process iHMMune-Align output.',
description='Process iHMMune-Align output.')
group_ihmm = parser_ihmm.add_argument_group('aligner parsing arguments')
group_ihmm.add_argument('-i', nargs='+', action='store', dest='aligner_files',
required=True,
help='''iHMMune-Align output file.''')
group_ihmm.add_argument('-r', nargs='+', action='store', dest='repo', required=True,
help='''List of folders and/or FASTA files containing
the set of germline sequences used by iHMMune-Align. These
reference sequences must contain IMGT-numbering spacers (gaps)
in the V segment.''')
group_ihmm.add_argument('-s', action='store', nargs='+', dest='seq_files',
required=True,
help='''List of input FASTA files (with .fasta, .fna or .fa
extension) containing sequences.''')
group_ihmm.add_argument('--10x', action='store', nargs='+', dest='cellranger_files',
help='''Table file containing 10X annotations (with .csv or .tsv
extension).''')
group_ihmm.add_argument('--asis-id', action='store_true', dest='asis_id',
help='''Specify to prevent input sequence headers from being parsed
to add new columns to database. Parsing of sequence headers requires
headers to be in the pRESTO annotation format, so this should be specified
when sequence headers are incompatible with the pRESTO annotation scheme.
Note, unrecognized header formats will default to this behavior.''')
group_ihmm.add_argument('--extended', action='store_true', dest='extended',
help='''Specify to include additional aligner specific fields in the output.
Adds the path score of the iHMMune-Align hidden Markov model as vdj_score;
adds fwr1, fwr2, fwr3, fwr4, cdr1, cdr2 and cdr3.''')
group_ihmm_validate = group_ihmm.add_mutually_exclusive_group(required=False)
# group_ihmm_validate.add_argument('--strict', action='store_const', const='strict', dest='validate',
# help='''By default, passing records must contain valid values for the
# V gene, J gene, junction region, and productivity call. If specified,
# this argument adds the additional requirement that the junction region must
# start at position 310 in the IMGT-numbered sequence.''')
group_ihmm_validate.add_argument('--partial', action='store_const', const='partial', dest='validate',
help='''If specified, include incomplete V(D)J alignments in
the pass file instead of the fail file. An incomplete alignment
is defined as a record that is missing a V gene assignment,
J gene assignment, junction region, or productivity call.''')
parser_ihmm.set_defaults(func=parseIHMM, validate='strict')
# Subparser to normalize AIRR file with IMGT-numbering
# desc_number = dedent('''
# Inserts IMGT numbering spacers into sequence_alignment, rebuilds the germline sequence
# in germline_alignment, and adjusts the values in the coordinate fields v_germline_start
# and v_germline_end accordingly.
# ''')
# parser_number = subparsers.add_parser('number', parents=[parser_parent],
# formatter_class=CommonHelpFormatter, add_help=False,
# help='Add IMGT-numbering to an AIRR Rearrangement TSV.',
# description=desc_number)
# group_number = parser_number.add_argument_group('aligner parsing arguments')
# group_number.add_argument('-i', nargs='+', action='store', dest='aligner_files', required=True,
# help='''AIRR Rearrangement TSV files.''')
# group_number.add_argument('-r', nargs='+', action='store', dest='repo', required=False,
# help='''List of folders and/or fasta files containing
# IMGT-numbered germline sequences corresponding to the
# set of germlines used for the alignment.''')
# parser_number.set_defaults(func=numberAIRR)
return parser
if __name__ == "__main__":
"""
Parses command line arguments and calls main
"""
parser = getArgParser()
checkArgs(parser)
args = parser.parse_args()
args_dict = parseCommonArgs(args, in_arg='aligner_files')
# Set no ID parsing if sequence files are not provided
if 'seq_files' in args_dict and not args_dict['seq_files']:
args_dict['asis_id'] = True
# Delete
if 'aligner_files' in args_dict: del args_dict['aligner_files']
if 'seq_files' in args_dict: del args_dict['seq_files']
if 'cellranger_files' in args_dict: del args_dict['cellranger_files']
if 'out_files' in args_dict: del args_dict['out_files']
if 'command' in args_dict: del args_dict['command']
if 'func' in args_dict: del args_dict['func']
# Call main
for i, f in enumerate(args.__dict__['aligner_files']):
args_dict['aligner_file'] = f
args_dict['out_file'] = args.__dict__['out_files'][i] \
if args.__dict__['out_files'] else None
if 'seq_files' in args.__dict__:
args_dict['seq_file'] = args.__dict__['seq_files'][i] \
if args.__dict__['seq_files'] else None
if 'cellranger_files' in args.__dict__:
args_dict['cellranger_file'] = args.__dict__['cellranger_files'][i] \
if args.__dict__['cellranger_files'] else None
args.func(**args_dict)
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