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#! /usr/bin/env python
# -*- coding: utf-8 -*-
"""
stand alone filter script based on jgi-rqc-pipeline/filter/fastq_filter2.py v3.2.8
Command
$ filter.py -f FASTQ.GZ -o OUT_DIR --skip-blast -html
Outputs
- normal filter outputs + index.html
Created: March 15, 2018
Shijie Yao (syao@lbl.gov)
Revision:
"""
import os
import sys
import argparse
import datetime
import shutil
import glob
import re
# import numpy as np
# import matplotlib
# matplotlib.use("Agg") ## This needs to skip the DISPLAY env var checking
# import matplotlib.pyplot as plt
# import mpld3
from pprint import pprint
## append the pipeline lib and tools relative path:
SRC_ROOT = os.path.abspath(os.path.dirname(os.path.abspath(__file__)))
sys.path.append(SRC_ROOT + "/lib") # common
import rqc_fastq as fastqUtil
from common import get_logger, get_status, checkpoint_step, set_colors, get_subsample_rate, append_rqc_file, append_rqc_stats
from os_utility import run_sh_command, make_dir_p
from rqc_utility import get_dict_obj, pipeline_val
from readqc import do_html_body as do_readqc_html_body
from html_utility import html_tag
## ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
## global vars
VERSION = "1.0.0"
SCRIPT_NAME = __file__
logLevel = "DEBUG"
DEBUG = False
color = {}
color = set_colors(color, True)
READQC_DIR = 'filtered-readqc'
PYDIR = os.path.abspath(os.path.dirname(__file__))
BBDIR = os.path.join(PYDIR, os.path.pardir)
FILTER_READ_COUNT = "read count"
FILTER_READ_SAMPLED_COUNT = "read sampled count"
STATUS_LOG_FNAME = 'status.log'
BB_STATS_LIST_FILE_NAME = 'filterStats.txt'
STATS_LIST_FILE_NAME = 'filter-stats.txt'
PIPE_START = 'start'
RQCFILTER_START = 'start rqcfilter'
RQCFILTER_END = 'end rqcfilter'
POST_START = 'start post process'
POST_END = 'end post process'
QC_START = 'start qc'
QC_END = 'end qc'
HTML_START = 'start html generation'
HTML_END = 'end html generation'
PIPE_COMPLETE = 'complete'
STEP_ORDER = {
PIPE_START : 0,
RQCFILTER_START : 10,
# rqcfilter.sh recorded steps:
'clumpify start' : 11,
'clumpify finish' : 12,
'ktrim start' : 20,
'ktrim finish' : 21,
'delete temp files start' : 30,
'delete temp files finish' : 31,
'filter start' : 40,
'filter finish' : 41,
'delete temp files start' : 50,
'delete temp files finish' : 51,
'short filter start' : 60,
'short filter finish' : 61,
'delete temp files start' : 70,
'delete temp files finish' : 71,
'removeCommonMicrobes start' : 80,
'removeCommonMicrobes finish' : 81,
'delete temp files start' : 90,
'delete temp files finish' : 91,
'dehumanize start' : 100,
'dehumanize finish' : 101,
'delete temp files start' : 110,
'delete temp files finish' : 111,
'merge start' : 120,
'merge finish' : 121,
'khist start' : 130,
'khist finish' : 131,
'rqcfilter complete' : 200,
RQCFILTER_END : 300,
POST_START : 381,
POST_END : 382,
QC_START : 400,
QC_END : 410,
HTML_START : 420,
HTML_END : 430,
PIPE_COMPLETE : 999
}
FILTER_METHODS_TXT = {"DNA": "dna.txt",
"FUNGAL": "fungal.txt",
"METAGENOME": "metagenome.txt",
"VIRAL-METAGENOME": "viral-metagenome.txt",
"ISO": "iso.txt",
"SAG": "sag.txt",
"CELL-ENRICHMENT": "cell-enrichment.txt",
"PLANT-2X150": "plant-2x150.txt",
"PLANT-2X250": "plant-2x250.txt",
"RNA": "rna.txt",
"RNAWOHUMAN": "rnawohuman.txt",
"3PRIMERNA": "3primerna.txt",
"SMRNA": "smrna.txt",
"METATRANSCRIPTOME": "mtaa.txt",
"MTF": "mtaa.txt",
"LFPE": "lfpe.txt",
"CLRS": "clrs.txt",
"CLIP-PE": "clip-pe.txt",
"NEXTERA-LMP": "nextera-lmp.txt",
"NEXTERA": "nextera-lmp.txt",
"NEXTSEQ": "nextseq.txt",
"ITAG": "itag.txt",
"MICROTRANS": "microtrans.txt",
"BISULPHITE": "bisulphite.txt",
"CHIPSEQ": "chip-seq.txt"}
## ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
## pipeline function definitions
def log_and_print(msg):
print(msg)
log.info(msg)
"""
Run rqcfilter.sh
@param fastq: the raw fastq input file [input of filtering]
@param outDir: the output directory
@param prodType: product type
@param status: current status
@return outFastqFile, outRrnaFastqFile
"""
def run_rqcfilter(infastq, outDir, prodType, status, enableRmoveMicrobes, enableAggressive, disableRmoveMicrobes, disableClumpify, taxList, rdb, log):
log_and_print("\n\n%s - RUN RQCFILTER <<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<%s\n" % (color['pink'], color['']))
make_dir_p(outDir)
opt = None
extraOptions = ""
optionFile = ""
if prodType.endswith("-OLD"):
extraOptions = " barcodefilter=f chastityfilter=f"
prodType = prodType.replace("-OLD", "")
if prodType in FILTER_METHODS_TXT:
optionFile = os.path.join(SRC_ROOT, "filter_param/", FILTER_METHODS_TXT[prodType].replace(".txt", ".config"))
log_and_print("Filter option file: %s" % optionFile)
opt = open(optionFile, 'r').readline().rstrip()
else:
log_and_print("The product type, %s, is not supported yet." % prodType)
sys.exit(2)
assert (opt), "Null filter options."
if prodType in ("METATRANSCRIPTOME", "MTF"):
if infastq.endswith(".gz"):
opt += " outribo=%s " % (os.path.basename(infastq).replace(".fastq", ".rRNA.fastq"))
else:
opt += " outribo=%s " % (os.path.basename(infastq).replace(".fastq", ".rRNA.fastq.gz"))
if enableRmoveMicrobes:
if opt.find("removemicrobes=f") != -1:
opt = opt.replace("removemicrobes=f", "removemicrobes=t")
opt += " removemicrobes=t "
if disableRmoveMicrobes:
if opt.find("removemicrobes=t") != -1:
opt = opt.replace("removemicrobes=t", "removemicrobes=f")
else: opt += " removemicrobes=f "
if enableAggressive:
opt += " aggressive=t microbebuild=3 "
if taxList:
opt += " taxlist=%s " % (taxList)
## Temp set clumpify=t for all prod types (RQC-890)
if not disableClumpify:
opt += " clumpify=t "
else:
opt += " clumpify=f "
opt += " tmpdir=null "
opt += extraOptions
cmd = os.path.join(BBDIR, "rqcfilter.sh")
filterLogFile = os.path.join(outDir, "filter.log")
cmdStr = "%s in=%s path=%s %s usejni=f rqcfilterdata=%s > %s 2>&1" % (cmd, infastq, outDir, opt, rdb, filterLogFile)
rtn = [None, status]
outFastqFile = None
shFileName = "%s/filter.sh" % outDir
def find_filtered_fastq(adir):
outFastqFile = None
searchPatt = os.path.join(adir, "*.fastq.gz")
outFastqFileList = glob.glob(searchPatt)
assert len(outFastqFileList) >= 1, "ERROR: cannot find *.fastq.gz output file."
for f in outFastqFileList:
f = os.path.basename(f)
t = f.split(".")
if t[-3] not in ("frag", "singleton", "unknown", "rRNA", "lmp"):
filterCode = t[-3]
elif t[-3] == "lmp": ## nextera
filterCode = t[-4]
if len(t) == 7: ## ex) 12345.1.1234.ACCCC.anqdpht.fastq.gz
fileNamePrefix = '.'.join(t[:4])
elif len(t) == 6: ## ex) 6176.5.39297.anqrpht.fastq.gz
fileNamePrefix = '.'.join(t[:3])
else:
log.warning("Unexpected filtered file name, %s", outFastqFileList)
fileNamePrefix = '.'.join(t[:-3])
log_and_print("Use %s as file prefix." %fileNamePrefix)
assert filterCode and fileNamePrefix, "ERROR: unexpected filter file name: %s" % (outFastqFileList)
of = os.path.join(adir, '.'.join([fileNamePrefix, filterCode, "fastq.gz"]))
lof = os.path.join(adir, '.'.join([fileNamePrefix, filterCode, "lmp.fastq.gz"]))
if os.path.isfile(of):
outFastqFile = of
elif os.path.isfile(lof):
outFastqFile = lof
else:
log.error("Cannot find fastq.gz file.")
# rename output file to *.filtered.fastq.gz
f = os.path.basename(outFastqFile)
t = f.split(".")
if t[-3] != 'filtered':
fto = os.path.join(adir, '.'.join(['.'.join(t[:-3]), 'filtered', "fastq.gz"]))
shutil.move(outFastqFile, fto)
outFastqFile = fto
return outFastqFile
def find_filter_number(outFastqFile):
filteredReadNum = fastqUtil.check_fastq_format(outFastqFile) / 4
if filteredReadNum < 0:
log_and_print("RUN RQCFILTER - filtered fastq format error: %s." % outFastqFile)
return filteredReadNum
if STEP_ORDER[status] < STEP_ORDER[RQCFILTER_END]:
create_shell(shFileName, (cmdStr,))
log_and_print("rqcfilter cmd=[%s]" % cmdStr)
log_and_print("sh file name=[%s]" % shFileName)
stdOut, stdErr, exitCode = run_sh_command(shFileName, True, log, True) ## stdOut of 0 is success
if exitCode != 0:
log.error("Failed to run : %s, stdout : %s, stderr: %s", shFileName, stdOut, stdErr)
return rtn
outFastqFile = find_filtered_fastq(outDir)
filteredReadNum = find_filter_number(outFastqFile)
log_and_print("Read counts after RQCFILTER step = %d" % filteredReadNum)
log_and_print("RUN RQCFILTER - completed")
checkpoint(RQCFILTER_END, status)
status = RQCFILTER_END
if filteredReadNum == 0:
log.warning("No reads left after filtering")
checkpoint(PIPE_COMPLETE, status)
with open(BB_STATS_LIST_FILE_NAME, 'a') as fh:
write_stats(fh, FILTER_READ_COUNT, 0, log)
write_stats(fh, FILTER_READ_BASE_COUNT, 0, log)
else:
log_and_print("No need to rerun RQCFILTER step, get filtered files and stats ... ")
outFastqFile = find_filtered_fastq(outDir)
rtn = [outFastqFile, status]
return rtn
"""
gzip the final filtered fastq file, and generate STATS_LIST_FILE_NAME log files.
@param fastq: the raw fastq file
@param outDir: output dir
@param filteredFastq: the filtered fastq file
@param status: where the pipeline was at by last run
@param log
@return filteredFastq or None if error
"""
def post_process(fastq, outDir, filteredFastq, status, log):
## obtain read counts from input and filtered fastq files and save the values to STATS_LIST_FILE_NAME file;
## compress the filtered fastq file
log_and_print("\n\n%s - RUN POST PROCESS <<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<%s\n" % (color['pink'], color['']))
if STEP_ORDER[status] < STEP_ORDER[POST_END]:
checkpoint(POST_START, status)
rawCnt = 0
rawBaseCnt = 0
newCnt = 0
newBaseCnt = 0
stats = get_dict_obj(BB_STATS_LIST_FILE_NAME)
rawCnt = pipeline_val('inputReads', {'type': 'int', 'vtype': 'numeric'}, stats)
rawBaseCnt = pipeline_val('inputBases', {'type': 'int', 'vtype': 'numeric'}, stats)
newCnt = pipeline_val('outputReads', {'type': 'int', 'vtype': 'numeric'}, stats)
newBaseCnt = pipeline_val('outputBases', {'type': 'int', 'vtype': 'numeric'}, stats)
readCounts = {}
readRmPct = 100.0 * ((rawCnt - newCnt) / float(rawCnt))
baseRmPct = 100.0 * ((rawBaseCnt - newBaseCnt) / float(rawBaseCnt))
readCounts['readRmPct'] = '%.3f' % readRmPct
readCounts['baseRmPct'] = '%.3f' % baseRmPct
refStats = {}
filterLogStat = {}
cardinality = None
bbdukVersion = None
bbmapVersion = None
if os.path.isfile("filter.log"):
with open(os.path.join(outDir, "filter.log"), "r") as FLFH:
isContamNumChecked = False ## Contamination will be done twice for removeribo or for MTF
isKtrimmedTotalRemovedNumChecked = False ## for parsing "Total Removed" after ktrimming
for l in FLFH:
if l.startswith("Input:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 2
if 'adaptertriminput' not in filterLogStat:
filterLogStat["adaptertriminput"] = {"numreads": toks[0], "numbases": toks[1]}
elif 'contamtriminput' not in filterLogStat:
filterLogStat["contamtriminput"] = {"numreads": toks[0], "numbases": toks[1]}
elif l.startswith("FTrimmed:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 4
filterLogStat["ftrimmed"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
elif l.startswith("KTrimmed:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 4
filterLogStat["ktrimmed"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
isKtrimmedTotalRemovedNumChecked = True
## RQCSUPPORT-1987
elif l.startswith("Total Removed:") and isKtrimmedTotalRemovedNumChecked:
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 4
filterLogStat["ktrimmed_total_removed"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
isKtrimmedTotalRemovedNumChecked = False
elif l.startswith("Trimmed by overlap:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 4
filterLogStat["trimmedbyoverlap"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
elif l.startswith("Result:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 4
if 'adaptertrimresult' not in filterLogStat:
filterLogStat["adaptertrimresult"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
elif 'contamtrimresult' not in filterLogStat:
filterLogStat["contamtrimresult"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
elif l.startswith("Unique 31-mers:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 2 or len(toks) == 1
if 'adaptertrimunique31mers' not in filterLogStat:
if len(toks) == 2:
filterLogStat["adaptertrimunique31mers"] = {"num": toks[1]}
else:
filterLogStat["adaptertrimunique31mers"] = {"num":"0"}
else:
if len(toks) == 2:
filterLogStat["contamtrimunique31mers"] = {"num": toks[1]}
else:
filterLogStat["contamtrimunique31mers"] = {"num":"0"}
elif not isContamNumChecked and l.startswith("Contaminants:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 4
filterLogStat["contaminants"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
isContamNumChecked = True
elif l.startswith("QTrimmed:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 4
filterLogStat["qtrimmed"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
elif l.startswith("Short Read Discards:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 4
filterLogStat["shortreaddiscards"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
elif l.startswith("Low quality discards:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 4
filterLogStat["lowqualitydiscards"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
elif l.startswith("BBDuk version"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 1
bbdukVersion = toks[0]
elif l.startswith("BBMap version"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 1
bbmapVersion = toks[0]
## BBDuk 36.12 06272016
elif l.startswith("Adapter Sequence Removed:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 4
filterLogStat["adaptersequenceremoved"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
elif l.startswith("Synthetic Contam Sequence Removed:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 4
filterLogStat["syntheticcontamsequenceremoved"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
## 08112016
elif l.startswith("Short Synthetic Contam Sequence Removed:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 4
filterLogStat["shortsyntheticcontamsequenceremoved"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
elif l.startswith("Ribosomal Sequence Removed:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 4
filterLogStat["ribosomalsequenceremoved"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
## BBMap 36.12 06272016
elif l.startswith("Human Sequence Removed:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 4
filterLogStat["humansequenceremoved"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
## RQC-862, RQC-880
elif l.startswith("Microbial Sequence Removed:"):
toks = re.findall("(\d+.\d*)", l.rstrip())
assert len(toks) == 4
filterLogStat["microbialremoved"] = {"numreads": toks[0], "percreads": toks[1], "numbases": toks[2], "percbases": toks[3]}
##
## refStats.txt format
##
## name %unambiguousReads unambiguousMB %ambiguousReads ambiguousMB unambiguousReads ambiguousReads
## human_masked 85.24693 498.92052 0.09378 0.55290 3350692 3686
## mouse_masked 0.03765 0.21670 0.10802 0.63690 1480 4246
## cat_masked 0.01862 0.09568 0.02514 0.14820 732 988
## dog_masked 0.00697 0.03815 0.01384 0.08160 274 544
##
if os.path.isfile("refStats.txt"):
refStatsFile = os.path.join(outDir, "refStats.txt")
with open(refStatsFile) as RFH:
## Need to report 0 if nothing matched
refStats['human'] = {"unambiguousReadsPerc":"0", "unambiguousMB":"0", "ambiguousReadsPerc":"0", "ambiguousMB":"0", "unambiguousReads":"0", "ambiguousReads":"0", "totalPerc":"0"}
refStats['cat'] = {"unambiguousReadsPerc":"0", "unambiguousMB":"0", "ambiguousReadsPerc":"0", "ambiguousMB":"0", "unambiguousReads":"0", "ambiguousReads":"0", "totalPerc":"0"}
refStats['dog'] = {"unambiguousReadsPerc":"0", "unambiguousMB":"0", "ambiguousReadsPerc":"0", "ambiguousMB":"0", "unambiguousReads":"0", "ambiguousReads":"0", "totalPerc":"0"}
refStats['mouse'] = {"unambiguousReadsPerc":"0", "unambiguousMB":"0", "ambiguousReadsPerc":"0", "ambiguousMB":"0", "unambiguousReads":"0", "ambiguousReads":"0", "totalPerc":"0"}
for l in RFH:
if l:
if l.startswith("#"):
continue
toks = l.rstrip().split()
assert len(toks) >= 7
## the number and percent of reads that map unambiguously or ambiguously to human, cat, dog.
## take the sum of the two numbers (ambiguous plus unambiguous) to use as the final percentage.
if l.startswith("human"):
refStats['human'] = {"unambiguousReadsPerc": toks[1], "unambiguousMB": toks[2], "ambiguousReadsPerc": toks[3], "ambiguousMB": toks[4], "unambiguousReads": toks[5], "ambiguousReads": toks[6], "totalPerc":float(toks[3])+float(toks[1])}
if l.startswith("cat"):
refStats['cat'] = {"unambiguousReadsPerc": toks[1], "unambiguousMB": toks[2], "ambiguousReadsPerc": toks[3], "ambiguousMB": toks[4], "unambiguousReads": toks[5], "ambiguousReads": toks[6], "totalPerc":float(toks[3])+float(toks[1])}
if l.startswith("dog"):
refStats['dog'] = {"unambiguousReadsPerc": toks[1], "unambiguousMB": toks[2], "ambiguousReadsPerc": toks[3], "ambiguousMB": toks[4], "unambiguousReads": toks[5], "ambiguousReads": toks[6], "totalPerc":float(toks[3])+float(toks[1])}
if l.startswith("mouse"):
refStats['mouse'] = {"unambiguousReadsPerc": toks[1], "unambiguousMB": toks[2], "ambiguousReadsPerc": toks[3], "ambiguousMB": toks[4], "unambiguousReads": toks[5], "ambiguousReads": toks[6], "totalPerc":float(toks[3])+float(toks[1])}
log.debug("refStats.txt: %s", str(refStats))
###########################################################
log_and_print("Write to stats file %s" % STATS_LIST_FILE_NAME)
###########################################################
if os.path.isfile(STATS_LIST_FILE_NAME):
os.remove(STATS_LIST_FILE_NAME)
with open(BB_STATS_LIST_FILE_NAME) as bbfh:
with open(STATS_LIST_FILE_NAME, 'a') as fh:
for line in bbfh:
if not line.startswith("#") and line.strip():
fh.write(line)
bbtoolsVersion = None
stats = get_dict_obj(STATS_LIST_FILE_NAME)
with open(STATS_LIST_FILE_NAME, 'a') as fh:
for key in readCounts:
if key not in stats:
write_stats(fh, key, readCounts[key], log)
for key in refStats:
for k in refStats[key]:
write_stats(fh, key+'_'+k, refStats[key][k], log)
write_stats(fh, "cardinality", cardinality, log)
## Write refStats to filterStats.txt file
for key in filterLogStat:
for k in filterLogStat[key]:
write_stats(fh, key + '_' + k, filterLogStat[key][k], log)
bbversionCmd = os.path.join(BBDIR, 'bbversion.sh')
cmd = "%s" % (bbversionCmd)
stdOut, _, exitCode = run_sh_command(cmd, True, log)
assert stdOut is not None
bbtoolsVersion = stdOut.strip()
## 05112017 Now bbtools version = bbmap version
# bbtoolsVersion = bbmapVersion if bbmapVersion else "37.xx"
assert bbtoolsVersion is not None
write_stats(fh, "filter_tool", "bbtools " + bbtoolsVersion, log)
write_stats(fh, "filter", VERSION, log)
## Version recording
if bbdukVersion is None: bbdukVersion = bbtoolsVersion
if bbmapVersion is None: bbmapVersion = bbtoolsVersion
write_stats(fh, "bbduk_version", bbdukVersion, log)
write_stats(fh, "bbmap_version", bbmapVersion, log)
checkpoint(POST_END, status)
status = POST_END
else:
log_and_print('No need to do post processing.')
return filteredFastq, status
##==============================================================================
## Helper functions
def clean_up(fList, log):
log_and_print("\n\n%s - CLEAN UP <<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<%s\n" % ( color['pink'], color['']))
for f in fList:
if os.path.isfile(f):
log_and_print("Removing %s ... ", f)
os.remove(f)
log_and_print("CLEAN UP - completed")
def create_shell(shName, cmdArray):
with open(shName, 'w') as fh:
fh.write("#!/bin/bash\n")
fh.write("set -e\n")
fh.write("set -o pipefail\n")
#fh.write("module unload blast+; module load blast+\n")
for item in cmdArray:
fh.write("%s\n" % item)
os.chmod(shName, 0755) #-rwxr-xr-x
## checkpoint logging
def checkpoint(status, fromStatus=PIPE_START):
if status == PIPE_START or STEP_ORDER[status] > STEP_ORDER[fromStatus]:
checkpoint_step(STATUS_LOG_FNAME, status)
def write_stats(fh, k, v, log):
line = "%s=%s" % (k, v)
fh.write("%s\n" % line)
def read_qc(odir, fastq, status):
log_and_print("\n\n%s - RUN QC <<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<%s\n" % (color['pink'], color['']))
if not os.path.isfile(fastq):
return False, status
qcdir = os.path.join(odir, READQC_DIR)
if STEP_ORDER[status] < STEP_ORDER[QC_END]:
checkpoint(QC_START, status)
qcexe = os.path.join(os.path.dirname(__file__), 'readqc.py')
cmd = '%s -o %s -f %s --skip-blast' % (qcexe, qcdir, fastq)
# print('DEBUG : %s' % cmd)
stdOut, stdErr, exitCode = run_sh_command(cmd, True)
if exitCode != 0:
print('ERROR : %s' % stdErr)
return False, qcdir, status
checkpoint(QC_END, status)
status = QC_END
else:
log_and_print("No need to do qc step.")
return True, qcdir, status
def do_html_body(odir, rawFastq, filteredFastq):
stats = get_dict_obj(os.path.join(odir, STATS_LIST_FILE_NAME))
tok_map = {
'inputReads' : {'token' : '[_RAW-READ-CNT_]', 'type': 'bigint'},
'inputBases' : {'token' : '[_RAW-BASE-CNT_]', 'type': 'bigint'},
'outputReads' : {'token' : '[_FILTERED-READ-CNT_]', 'type': 'bigint'},
'outputBases' : {'token' : '[_FILTERED-BASE-CNT_]', 'type': 'bigint'},
'readRmPct' : {'token' : '[_REMOVED-READ-PCT_]', 'type': 'raw'},
'baseRmPct' : {'token' : '[_REMOVED-BASE-PCT_]', 'type': 'raw'},
'lowqualitydiscards_numreads' : {'token' : '[_LOW-QUAL-REMOVED-READ-CNT_]', 'type': 'bigint', 'filter': 0},
'lowqualitydiscards_percreads' : {'token' : '[_LOW-QUAL-REMOVED-PCT_]', 'type': 'raw', 'filter': 0},
'contaminants_numreads' : {'token' : '[_ARTI-REMOVED-READ-CNT_]', 'type': 'bigint', 'filter': 0},
'contaminants_percreads' : {'token' : '[_ARTI-REMOVED-PCT_]', 'type': 'raw', 'filter': 0},
'ribosomalsequenceremoved_numreads' : {'token' : '[_RRNA-REMOVED-READ-CNT_]', 'type': 'bigint', 'filter': 0},
'ribosomalsequenceremoved_percreads' : {'token' : '[_RRNA-REMOVED-READ-PCT_]', 'type': 'raw', 'filter': 0},
'microbialremoved_numreads' : {'token' : '[_MICROBE-REMOVED-READ-CNT_]', 'type': 'bigint', 'filter': 0},
'microbialremoved_percreads' : {'token' : '[_MICROBE-REMOVED-READ-PCT_]', 'type': 'raw', 'filter': 0},
'human_unambiguousreads' : {'token' : '[_HUMAN-REMOVED-READ-CNT_]', 'type': 'bigint', 'filter': 0},
'human_unambiguousreadsperc' : {'token' : '[_HUMAN-REMOVED-READ-PCT_]', 'type': 'raw', 'filter': 0},
'dog_unambiguousreads' : {'token' : '[_DOG-REMOVED-READ-CNT_]', 'type': 'bigint', 'filter': 0},
'dog_unambiguousreadsperc' : {'token' : '[_DOG-REMOVED-READ-PCT_]', 'type': 'raw', 'filter': 0},
'cat_unambiguousreads' : {'token' : '[_CAT-REMOVED-READ-CNT_]', 'type': 'bigint', 'filter': 0},
'cat_unambiguousreadsperc' : {'token' : '[_CAT-REMOVED-READ-PCT_]', 'type': 'raw', 'filter': 0},
'mouse_unambiguousreads' : {'token' : '[_MOUSE-REMOVED-READ-CNT_]', 'type': 'bigint', 'filter': 0},
'mouse_unambiguousreadsperc' : {'token' : '[_MOUSE-REMOVED-READ-PCT_]', 'type': 'raw', 'filter': 0},
}
temp = os.path.join(PYDIR, 'template/filter_body_template.html')
html = ''
with open(temp, 'r') as fh:
html = fh.read()
## do the place-holder replacement !!
html = html.replace('[_RAW-FILE-LOCATION_]', rawFastq)
html = html.replace('[_FILTERED-FILE-LOCATION_]', filteredFastq)
fsize = format(os.stat(rawFastq).st_size / (1024*1024), ',')
html = html.replace('[_RAW-FILE-SIZE_]', fsize)
fsize = format(os.stat(filteredFastq).st_size / (1024*1024), ',')
html = html.replace('[_FILTERED-FILE-SIZE_]', fsize)
for key in tok_map:
dat = tok_map[key]
html = html.replace(dat['token'], pipeline_val(key, dat, stats))
# readqc on the filter file
if qcdir:
hbody = do_readqc_html_body(qcdir, odir)
else:
hbody = ''
html = html.replace('[_FILTERED-READ-QC_]', hbody)
return html
def do_html(odir, qcdir, rawFastq, filteredFastq, status):
log_and_print("\n\n%s - Create HTML file <<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<<%s\n" % (color['pink'], color['']))
fname = os.path.basename(rawFastq)
stats = get_dict_obj(os.path.join(odir, STATS_LIST_FILE_NAME))
temp = os.path.join(PYDIR, 'template/template.html')
with open(temp, 'r') as fh:
html = fh.read()
html = html.replace('[_PAGE-TITLE_]', 'Filter Report')
html = html.replace('[_REPORT-TITLE_]', 'BBTools Filtering Report')
html = html.replace('[_INPUT-FILE-NAME_]', fname)
html = html.replace('[_REPORT-DATE_]', '{:%Y-%m-%d %H:%M:%S}'.format(datetime.datetime.now()))
hbody = do_html_body(odir, rawFastq, filteredFastq)
html = html.replace('[_REPORT-BODY_]', hbody)
fbasename = 'filter.log'
fname = os.path.join(outputPath, fbasename)
html = html.replace('[_FILTER-LOG_]', html_tag('a', fbasename, {'href': fbasename}))
fsize = '%.1f' % (float(os.stat(fname).st_size) / 2014.0)
html = html.replace('[_FILTER-LOG-SIZE_]', fsize)
# fbasename = 'filter.txt'
# fname = os.path.join(outputPath, fbasename)
# html = html.replace('[_FILTER-REPORT_]', html_tag('a', fbasename, {'href': fbasename}))
# fsize = '%.1f' % (float(os.stat(fname).st_size) / 2014.0)
# html = html.replace('[_FILTER-REPORT-SIZE_]', fsize)
## write the html to file
idxfile = os.path.join(odir, 'index.html')
with open(idxfile, 'w') as fh2:
fh2.write(html)
print('HTML index file written to %s' % idxfile)
# copy the css file
cssdir = os.path.join(PYDIR, 'css')
todir = os.path.join(odir, 'css')
if os.path.isdir(todir):
shutil.rmtree(todir)
shutil.copytree(cssdir, todir, False, None)
# copy the image file
imgdir = os.path.join(PYDIR, 'images')
todir = os.path.join(odir, 'images')
if os.path.isdir(todir):
shutil.rmtree(todir)
shutil.copytree(imgdir, todir, False, None)
return status
## ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
## main program
if __name__ == "__main__":
## Parse options
usage = "* Filter Pipeline, version %s\n" % (VERSION)
origCmd = ' '.join(sys.argv)
## command line options
parser = argparse.ArgumentParser(description=usage, formatter_class=argparse.ArgumentDefaultsHelpFormatter)
PROD_TYPE = [
'DNA',
'FUNGAL',
'METAGENOME',
'VIRAL-METAGENOME',
'SAG',
'ISO',
'SAG',
'CELL-ENRICHMENT',
'PLANT-2x150',
'PLANT-2x250',
'RNA',
'SMRNA',
'METATRANSCRIPTOME',
'LFPE',
'CLRS',
'CLIP-PE',
'NEXTERA',
'ITAG',
'MICROTRANS',
'BISULPHITE',
'3PRIMERNA',
'CHIPSEQ',
'RNAWOHUMAN'
]
parser.add_argument("-f", "--fastq", dest="fastq", help="Set input fastq file (full path to fastq)", required=True)
parser.add_argument("-o", "--output-path", dest="outputPath", help="Set output path to write to", required=True)
parser.add_argument("-p", "--prod-type", dest="prodType", help="Set product type: %s" % PROD_TYPE, required=True)
parser.add_argument("-rdb", "--ref-databases", dest="rqcfilterdata", help="Path to RQCFilterData dir", required=True)
parser.add_argument("-t", "--taxlist", dest="taxList", help="A list of taxid(s) to exclude in CSV format", required=False)
parser.add_argument("-ap", "--ap", dest="apNum", help="Set AP (Analysis Project) ID. Ex) -ap 123 or -ap 123,456,789", required=False)
parser.add_argument("-at", "--at", dest="atNum", help="Set AT (Analysis Task) ID. Ex) -at 123 or -at 123,456,789", required=False)
parser.add_argument("-v", "--version", action="version", version=VERSION)
## switches
parser.add_argument("-qc", "--qc", dest="doqc", action="store_true", help="also perform readqc analysis on the filtered output", default=False)
parser.add_argument("-a", "--aggressive", dest="enableAggressive", action="store_true", help="Enable aggressive=t and microbebuild=3", default=False)
parser.add_argument("-b", "--skip-blast", dest="skipBlastFlag", action="store_true", help="Skip Blast search", default=False)
parser.add_argument("-c", "--skip-cleanup", dest="skipCleanUp", action="store_true", help="Skip file clean up after pipeline complete", default=False)
parser.add_argument("-d", "--debug", dest="doDebug", action="store_true", help="Enable debug mode")
parser.add_argument("-l", "--disable-clumpify", dest="disableClumpify", action="store_true", help="Disable clumpify", default=False)
parser.add_argument("-m", "--skip-microbes-removal", dest="disableRmoveMicrobes", action="store_true", help="Skip microbes removal", default=False)
parser.add_argument("-r", "--contam", dest="enableRmoveMicrobes", action="store_true", help="Enable removemicrobes=t", default=False)
parser.add_argument("-s", "--skip-subsampleqc", dest="skipSubsampleQc", action="store_true", help="Skip the subsample and qc step", default=False)
parser.add_argument("-pl", "--print-log", dest="print_log", default = False, action = "store_true", help = "print log to screen")
## produce html when processing is done
parser.add_argument("-html", "--html", action="store_true", help="Create html file", dest="html", default=False, required=False)
skipCleanUp = False
skipSubsampleQc = False
outputPath = None ## output path, defaults to current working directory
fastq = None ## full path to input fastq
# logLevel = "DEBUG"
enableRmoveMicrobes = False
disableRmoveMicrobes = False
disableClumpify = False
enableAggressive = False
skipBlastFlag = False
apNum = None
atNum = None
taxList = ""
options = parser.parse_args()
print_log = options.print_log
if options.outputPath:
outputPath = options.outputPath
if not outputPath:
outputPath = os.getcwd()
## create output_directory if it doesn't exist
if not os.path.isdir(outputPath):
os.makedirs(outputPath)
outputPath = os.path.realpath(outputPath)
outputPath = outputPath.replace("/chos", "") if outputPath.startswith("/chos") else outputPath
## initialize my logger
logFile = os.path.join(outputPath, "rqc_filter_pipeline.log")
print "Started filtering pipeline with %s, writing log to: %s" % (SCRIPT_NAME, logFile)
log = get_logger("filter", logFile, logLevel, print_log, True)
if options.doDebug:
DEBUG = True
if options.skipCleanUp:
skipCleanUp = True
if options.skipBlastFlag:
skipBlastFlag = True
if options.skipSubsampleQc:
skipSubsampleQc = True
if options.enableRmoveMicrobes:
if options.disableRmoveMicrobes:
log.error("Conflict in option parameters: cannot set skip-contam with skip-microbes-removal.")
sys.exit(0)
enableRmoveMicrobes = True
if options.disableRmoveMicrobes:
if options.enableRmoveMicrobes:
log.error("Conflict in option parameters: cannot set skip-contam with skip-microbes-removal.")
sys.exit(0)
disableRmoveMicrobes = True
if options.enableAggressive:
enableAggressive = True
if options.disableClumpify:
disableClumpify = True
if options.fastq:
fastq = options.fastq
if options.taxList:
taxList = options.taxList
if options.apNum:
apNum = options.apNum
if options.atNum:
atNum = options.atNum
log_and_print("%s" % '#' * 80)
log_and_print(" Filtering pipeline (version %s)" % VERSION)
log_and_print("%s" % '#' * 80)
prodType = options.prodType.upper()
skipSubsampleQcFlag = options.skipSubsampleQc ## run subsample and qc or not
if not os.path.isdir(outputPath):
log.error("Cannot work with directory: %s", outputPath)
## check for fastq file
if fastq:
if not os.path.isfile(fastq):
log.error("Input fastq file, %s not found, abort!", fastq)
else:
log.error("No fastq defined, abort!")
fastq = os.path.realpath(fastq)
fastq = fastq.replace("/chos", "") if fastq.startswith("/chos") else fastq
##--------------------------------
## init log
log_and_print("Started pipeline, writing log to: %s" % logFile)
log_and_print("CMD: %s" % origCmd)
log_and_print("Fastq file: %s" % fastq)
log_and_print("Output path: %s" % outputPath)
os.chdir(outputPath)
cycle = 0
cycleMax = 1
bIsPaired = False
status = get_status(STATUS_LOG_FNAME, log)
log_and_print("Starting pipeline at [%s]" % status)
if status == PIPE_START:
checkpoint(PIPE_START)
## main loop: retry upto cycleMax times
while cycle < cycleMax:
cycle += 1
log_and_print("ATTEMPT [%d]" % cycle)
filesToRemove = [] # list of intermediate files for clean up
lastFastq = fastq # lastFastq : fastq produced by each step, init to input
rRnaFilterFile = None # MTF generates this 2nd filtered output file
subsampledFastq = None
bIsPaired = None
filteredReadNum = -1
filteredReadNumRrna = -1
FragFile = SingletonFile = UnknownFile = None
if cycle > 1:
status = get_status(STATUS_LOG_FNAME, log)
if status != PIPE_COMPLETE:
##
## Run rqcfilter.sh
##
lastFastq, status = run_rqcfilter(lastFastq, outputPath, prodType, status, enableRmoveMicrobes, enableAggressive, disableRmoveMicrobes, disableClumpify, taxList, options.rqcfilterdata, log) ## Only MTF type generates rRnaFilterFile
if filteredReadNum == 0:
break
##--------------------------------
## Run post processing
if lastFastq is not None and lastFastq != -1:
lastFastq, status = post_process(fastq, outputPath, lastFastq, status, log)
else:
print "Failed @ rqcfilter"
##--------------------------------
## Clean up
if lastFastq is not None and lastFastq != -1:
## run readQC on the filtered fastq
if options.doqc:
rtn, qcdir, status = read_qc(outputPath, lastFastq, status)
else:
qcdir = None
rtn = True
## create html file on the readQC results
if rtn:
status = do_html(outputPath, qcdir, fastq, lastFastq, status)
checkpoint(PIPE_COMPLETE)
if not skipCleanUp:
clean_up(filesToRemove, log)
else:
log_and_print("SKIP CLEANUP")
log_and_print("Pipeline Completed")
cycle = cycleMax + 1
else:
print "Failed @ postprocess"
else:
cycle = cycleMax + 1
log_and_print("Pipeline already completed")
print "Done."
sys.exit(0)
## EOF
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