Mercurial > repos > iss > eurl_vtec_wgs_pt

annotate EURL_VTEC_WGS_PT.py @ 0:c6bab5103a14 draft

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"planemo upload commit 6abf3e299d82d07e6c3cf8642bdea80e96df64c3-dirty"
author iss
date Mon, 21 Mar 2022 15:23:09 +0000
parents
children 444b0421bbdc
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0
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1 #!/usr/bin/env python3
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2 # -*- coding: utf-8 -*-
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3 """
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4 ############################################################################
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5 # Istituto Superiore di Sanita'
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6 # European Union Reference Laboratory (EU-RL) for Escherichia coli, including Verotoxigenic E. coli (VTEC)
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7 # Developer: Arnold Knijn arnold.knijn@iss.it
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8 ############################################################################
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9 """
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10
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11 import argparse
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12 import sys
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13 import os
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14 import shutil
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15 import subprocess
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16 import HTML
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17 import datetime
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18 import fileinput
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19
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20 BASE_URL = 'https://aries.iss.it'
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21 TOOL_DIR = os.path.dirname(os.path.abspath(__file__))
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22
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23 def insertFile(filename, report):
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24 with open(filename) as html_in:
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25 for line in html_in:
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26 report.write(line)
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27
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28 def insertFileAsTable(filename, report, hasheader=False, tabclass="table table-rep"):
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29 with open(filename) as table_in:
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30 table_data = [[str(col) for col in row.split('\t')] for row in table_in]
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31 insertTable(table_data, report, hasheader, tabclass)
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32
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33 def insertTable(table_data, report, hasheader=False, tabclass="table table-rep"):
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34 if hasheader:
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35 htmlcode = HTML.table(table_data[1:], attribs={'class':tabclass}, header_row=table_data[0])
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36 else:
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37 htmlcode = HTML.table(table_data, attribs={'class':tabclass})
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38 report.write(htmlcode)
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39
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40 def openFileAsTable(filename):
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41 with open(filename) as table_in:
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42 table_data = [[str(col).rstrip() for col in row.split('\t')] for row in table_in]
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43 return table_data
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44
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45 def __main__():
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46 parser = argparse.ArgumentParser()
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47 parser.add_argument('--serotyping', dest='serotyping', help='perform serotyping', action='store_true')
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48 parser.add_argument('--virulotyping', dest='virulotyping', help='perform virulotyping', action='store_true')
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49 parser.add_argument('--shigatoxintyping', dest='shigatoxintyping', help='perform shigatoxintyping', action='store_true')
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50 parser.add_argument('--amrtyping', dest='amrtyping', help='perform amrtyping', action='store_true')
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51 parser.add_argument('-1', '--input1', dest='input1', help='forward or single-end reads file in Sanger FASTQ format')
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52 parser.add_argument('--input1_ext', dest='input1_ext', help='extension of forward or single-end reads file in Sanger FASTQ format')
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53 parser.add_argument('--input1_name', dest='input1_name', help='name of forward or single-end reads file in Sanger FASTQ format')
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54 parser.add_argument('-2', '--input2', dest='input2', help='reverse reads file in Sanger FASTQ format')
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55 parser.add_argument('--input2_ext', dest='input2_ext', help='extension of reverse reads file in Sanger FASTQ format')
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56 parser.add_argument('--input2_name', dest='input2_name', help='name of reverse reads file in Sanger FASTQ format')
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57 parser.add_argument('--html1', dest='html1', help='html FASTQC file')
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58 parser.add_argument('--html1_id', dest='html1_id', help='html FASTQC file id')
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59 parser.add_argument('--html1_path', dest='html1_path', help='html FASTQC file path')
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60 parser.add_argument('--text1', dest='text1', help='text FASTQC file')
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61 parser.add_argument('--html2', dest='html2', help='html FASTQC file')
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62 parser.add_argument('--html2_id', dest='html2_id', help='html FASTQC file id')
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63 parser.add_argument('--html2_path', dest='html2_path', help='html FASTQC file path')
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64 parser.add_argument('--text2', dest='text2', help='text FASTQC file')
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65 parser.add_argument('--contigs', dest='contigs', help='Assembly contigs')
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66 parser.add_argument('--quast', dest='quast', help='Quast report')
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67 parser.add_argument('--log', dest='logfile', help='log file')
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68 parser.add_argument('--virulotyper', dest='virulotyper', help='Virulotyping Mapping reads')
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69 parser.add_argument('--virulotyper_id', dest='virulotyper_id', help='Virulotyping Mapping reads id')
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70 parser.add_argument('--stx', dest='stx', help='Shiga toxin')
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71 parser.add_argument('--mlstsevenloci', dest='mlstsevenloci', help='Multi Locus Alleles table')
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72 parser.add_argument('--amr', dest='amrgenes', help='AMR genes')
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73 parser.add_argument('--amr_id', dest='amr_id', help='AMR file id')
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74 parser.add_argument('--antigen_O', dest='antigen_O', help='Antigen for O')
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75 parser.add_argument('--antigen_H', dest='antigen_H', help='Antigen for H')
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76 parser.add_argument('--output', dest='output', help='output report html file')
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77 args = parser.parse_args()
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78
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79 log = open(args.logfile, 'w')
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80 log.write("EURL VTEC WGS PT v3.2\n\nTool versions\n=============\n")
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81 os.system("ln -s $(readlink -e $(which trimmomatic)).jar trimmomatic.jar")
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82 # FASTQC
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83 subprocess.call("python " + TOOL_DIR + "/scripts/rgFastQC.py -i " + args.input1 + " -d " + args.html1_path + " -o " + args.html1 + " -t " + args.text1 + " -f " + args.input1_ext + " -j " + args.input1_name + " -e " + "fastqc", shell=True)
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84 log.write(os.popen("fastqc -v").read())
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85 if args.input2:
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86 # FASTQC
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87 subprocess.call("python " + TOOL_DIR + "/scripts/rgFastQC.py -i " + args.input2 + " -d " + args.html2_path + " -o " + args.html2 + " -t " + args.text2 + " -f " + args.input2_ext + " -j " + args.input2_name + " -e " + "fastqc", shell=True)
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88 # TRIMMING
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89 subprocess.call("java ${_JAVA_OPTIONS:--Xmx8G} -jar trimmomatic.jar PE -threads ${GALAXY_SLOTS:-6} -phred33 " + args.input1 + " " + args.input2 + " trimmed1.fq trimmed1unpaired trimmed2.fq trimmed2unpaired SLIDINGWINDOW:5:20 LEADING:3 TRAILING:3 MINLEN:36", shell=True)
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90 log.write("\nTrimmomatic v0.39\n")
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91 log.write("parameters: phred33 SLIDINGWINDOW:5:20 LEADING:3 TRAILING:3 MINLEN:36\n\n")
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92 # ASSEMBLY
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93 subprocess.call("perl " + TOOL_DIR + "/scripts/spades.pl spades_contigs spades_contig_stats spades_scaffolds spades_scaffold_stats spades_log NODE spades.py --disable-gzip-output --isolate -t ${GALAXY_SLOTS:-16} --pe1-ff --pe1-1 trimmed1.fq --pe1-2 trimmed2.fq", shell=True)
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94 subprocess.call("perl " + TOOL_DIR + "/scripts/filter_spades_repeats.pl -i spades_contigs -t spades_contig_stats -c 0.33 -r 1.75 -l 1000 -o output_with_repeats -u output_without_repeats -n repeat_sequences_only -e 5000 -f discarded_sequences -s summary", shell=True)
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95 shutil.move("output_without_repeats", args.contigs)
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96 log.write(os.popen("spades.py -v").read())
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97 log.write("parameters: --isolate, pe1-ff, pe1-1, pe1-2 filter_repeats\n\n")
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98 else:
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99 # TRIMMING
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100 subprocess.call("java ${_JAVA_OPTIONS:--Xmx8G} -jar trimmomatic.jar SE -threads ${GALAXY_SLOTS:-6} -phred33 " + args.input1 + " trimmed1.fq SLIDINGWINDOW:5:20 LEADING:3 TRAILING:3 MINLEN:55", shell=True)
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101 log.write("\nTrimmomatic v0.39\n")
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102 log.write("parameters: phred33 SLIDINGWINDOW:5:20 LEADING:3 TRAILING:3 MINLEN:55\n\n")
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103 # ASSEMBLY
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104 subprocess.call("perl " + TOOL_DIR + "/scripts/spades.pl spades_contigs spades_contig_stats spades_scaffolds spades_scaffold_stats spades_log NODE spades.py --disable-gzip-output --isolate -t ${GALAXY_SLOTS:-16} --iontorrent -s trimmed1.fq", shell=True)
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105 subprocess.call("perl " + TOOL_DIR + "/scripts/filter_spades_repeats.pl -i spades_contigs -t spades_contig_stats -c 0.33 -r 1.75 -l 1000 -o output_with_repeats -u output_without_repeats -n repeat_sequences_only -e 5000 -f discarded_sequences -s summary", shell=True)
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106 shutil.move("output_without_repeats", args.contigs)
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107 log.write(os.popen("spades.py -v").read())
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108 log.write("parameters: --isolate, --iontorrent filter_repeats\n\n")
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109 # QUAST
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110 subprocess.call("quast --threads 4 -o outputdir --est-ref-size 5000000 --min-contig 500 -l '" + args.input1_name + "' --contig-thresholds 0,1000 " + args.contigs, shell=True)
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111 shutil.move("outputdir/report.tsv", args.quast)
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112 if args.virulotyping:
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113 # VIRULOTYPER
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114 if args.input2:
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115 subprocess.call("perl " + TOOL_DIR + "/scripts/patho_typing.pl 'python " + TOOL_DIR + "/scripts/patho_typing.py -s Escherichia coli -f " + args.input1 + " " + args.input2 + " -o output_dir -j 4 --minGeneCoverage 90 --minGeneIdentity 90 --minGeneDepth 15'", shell=True)
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116 else:
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117 subprocess.call("perl " + TOOL_DIR + "/scripts/patho_typing.pl 'python " + TOOL_DIR + "/scripts/patho_typing.py -s Escherichia coli -f " + args.input1 + " -o output_dir -j 4 --minGeneCoverage 90 --minGeneIdentity 90 --minGeneDepth 15'", shell=True)
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118 subprocess.call("(head -n 1 pathotyper_rep_tot_tab && tail -n +2 pathotyper_rep_tot_tab | sort -k 2rn) > " + args.virulotyper, shell=True)
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119 log.write("\n\nViruloTyper\n===========\npatho_typing v1.0\n")
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120 log.write("parameters: minGeneCoverage=90, minGeneIdentity=90, minGeneDepth=15\n\n")
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121 log.write(os.popen("cat " + TOOL_DIR + "/data/ViruloTyping_db.txt").read())
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122 # SEQUENCETYPER
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123 subprocess.call("mlst --legacy --scheme ecoli " + args.contigs + " | cut -f3,4,5,6,7,8,9,10 > " + args.mlstsevenloci, shell=True)
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124 sequence_typing = openFileAsTable(args.mlstsevenloci)
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125 log.write("\n\nSequence Typer\n==============\n")
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126 log.write(os.popen("mlst -v").read())
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127 log.write("\n")
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128 log.write(os.popen("cat " + TOOL_DIR + "/data/SequenceTyping_db.txt").read())
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129 if args.shigatoxintyping:
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130 # SHIGATOXIN TYPER
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131 if args.input2:
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132 # CONSENSUS
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133 subprocess.call("sh " + TOOL_DIR + "/scripts/stx_subtype_pe.sh " + TOOL_DIR + " trimmed1.fq trimmed2.fq " + args.contigs, shell=True)
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134 else:
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135 # CONSENSUS
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136 subprocess.call("sh " + TOOL_DIR + "/scripts/stx_subtype_se.sh " + TOOL_DIR + " trimmed1.fq " + args.contigs, shell=True)
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137 # SHIGATOXIN SEQUENCE SEARCH
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138 subprocess.call("sh " + TOOL_DIR + "/scripts/stx_subtype_fa.sh " + TOOL_DIR + " stx.fasta", shell=True)
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139 subprocess.call("echo 'sseqid\tpident\tlength\tpositive' > shigatoxin_fct", shell=True)
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140 subprocess.call("cat shigatoxin_fc >> shigatoxin_fct", shell=True)
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141 shutil.move("shigatoxin_fct", args.stx)
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142 shigatoxin_typing = openFileAsTable("shigatoxin_fc")
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143 log.write("\n\nShigatoxin Typer v2.0\n==============\n")
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144 log.write(os.popen("cat " + TOOL_DIR + "/data/ShigatoxinTyping_db.txt").read())
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145 if args.serotyping:
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146 # SEROTYPER
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147 subprocess.call("echo 'sseqid\tpident\tlength\tpositive' > serogroup_OH_fcd", shell=True)
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148 if args.input2:
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149 subprocess.call("sh " + TOOL_DIR + "/scripts/serotype.sh " + TOOL_DIR + " y " + args.input1 + " " + args.input2 + " " + args.contigs, shell=True)
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150 else:
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151 subprocess.call("sh " + TOOL_DIR + "/scripts/serotype.sh " + TOOL_DIR + " n " + args.input1 + " xxx " + args.contigs, shell=True)
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152 # SEROTYPER O
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153 subprocess.call("awk -F '\t' '$4>800 { print $2 FS $3 FS $4 FS $16 }' serogroup_O | sort -nrk 2 -nrk 3 > serogroup_O_fc", shell=True)
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154 subprocess.call("awk -F , '!seen[$0]++' serogroup_O_fc > serogroup_O_fcd", shell=True)
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155 sero_typing_o = openFileAsTable("serogroup_O_fcd")
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156 subprocess.call("cat serogroup_O_fcd >> serogroup_OH_fcd", shell=True)
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157 shutil.move("serogroup_O_fcd", args.antigen_O)
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158 # SEROTYPER H
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159 subprocess.call("awk -F '\t' '$4>800 { print $2 FS $3 FS $4 FS $16 }' serogroup_H | sort -nrk 2 -nrk 3 > serogroup_H_fc", shell=True)
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160 subprocess.call("awk -F , '!seen[$0]++' serogroup_H_fc > serogroup_H_fcd", shell=True)
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161 sero_typing_h = openFileAsTable("serogroup_H_fcd")
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162 subprocess.call("cat serogroup_H_fcd >> serogroup_OH_fcd", shell=True)
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163 shutil.move("serogroup_H_fcd", args.antigen_H)
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164 if os.stat(args.antigen_O).st_size == 0 and os.stat(args.antigen_H).st_size == 0:
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165 subprocess.call("echo '-\t-\t-\t-' >> serogroup_OH_fcd", shell=True)
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166 log.write("\n\nSero Typer\n==============\n")
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167 log.write(os.popen("cat " + TOOL_DIR + "/data/SeroTyping_db.txt").read())
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168 if args.amrtyping:
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169 # AMRGENES
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170 # subprocess.call("amrfinder --threads 4 --database " + TOOL_DIR + "/data/amrfinder -n " + args.contigs + " -O Escherichia -o " + args.amrgenes, shell=True)
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171 subprocess.call("abricate --db resfinder " + args.contigs + " > " + args.amrgenes, shell=True)
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172 log.write("\n\nAMR Typer\n==============\nabricate ")
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173 log.write(os.popen("abricate --version").read())
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174 log.write("\ndatabase version: ")
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175 log.write(os.popen("abricate --list | grep resfinder").read())
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176 # REPORT
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177 try:
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178 report = open(args.output, 'w')
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179 # write head html
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180 insertFile(TOOL_DIR + "/report_head.html", report)
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181 report.write("<td><h1>EURL VTEC WGS PT</h1><h2>Report for %s</h2>%s</td>" % (args.input1_name, datetime.datetime.utcnow().strftime("%Y-%m-%d %H:%M UTC")))
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182 insertFile(TOOL_DIR + "/report_head2.html", report)
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183 # write results
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184 report.write("<h3>Summary</h3>\n")
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185 if args.serotyping:
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186 report.write("<p>Serotype: ")
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187 if len(sero_typing_o) == 0:
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188 report.write("O?")
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189 else:
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190 report.write("%s" % sero_typing_o[0][0][sero_typing_o[0][0].rfind("O"):])
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191 if len(sero_typing_h) == 0:
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192 report.write(":H?")
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193 else:
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194 report.write(":%s" % sero_typing_h[0][0][sero_typing_h[0][0].rfind("H"):])
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195 report.write("</p>\n")
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196 report.write("<p>Sequence type: ")
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197 if len(sequence_typing) < 2:
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198 report.write("Sequence typing failed")
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199 elif sequence_typing[1][1] == "-":
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200 report.write("Sequence typing failed")
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201 else:
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202 report.write("ST%s" % sequence_typing[1][0])
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203 report.write("</p>\n")
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204 if args.virulotyping:
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205 subprocess.call("sort " + args.virulotyper + " | awk '/eae_|stx1._|stx2._|ehxa_/ && $2>50 && !seen[substr($1, 1, index($1, \"_\")-2)]++ { printf(\"%s%s\",sep,substr($1, 1, index($1, \"_\")-1));sep=\", \" }END{print \"\"}' > virulotyper_rep", shell=True)
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206 for line in fileinput.input("virulotyper_rep", inplace=True):
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207 print(line.replace("1a", "1"),)
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208 for line in fileinput.input("virulotyper_rep", inplace=True):
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209 print(line.replace("2a", "2"),)
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210 for line in fileinput.input("virulotyper_rep", inplace=True):
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211 print(line.replace("1b", "1"),)
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212 for line in fileinput.input("virulotyper_rep", inplace=True):
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213 print(line.replace("2b", "2"),)
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214 report.write("<p>Virulotypes: ")
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215 insertFile("virulotyper_rep", report)
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216 report.write("</p>\n")
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217 if args.shigatoxintyping:
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218 report.write("<p>Stx Subtypes: ")
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219 if len(shigatoxin_typing) == 0:
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220 str_shigatoxin_subtype = "No subtype match found"
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221 else:
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222 # get corresponding subtypes
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223 str_shigatoxin_subtype = ""
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224 shigatoxin_subtypes = []
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225 shigatoxin_subtypes_raw = []
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226 shigatoxin_types = openFileAsTable(TOOL_DIR + "/data/stx_subtypes")
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227 for subtype in shigatoxin_typing:
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228 blast_pident_100 = float(subtype[1]) == 100
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229 if (blast_pident_100):
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230 for item in shigatoxin_types:
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231 if item[0] == subtype[0] and item[1] not in shigatoxin_subtypes_raw:
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232 shigatoxin_subtypes.append(item[1])
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233 shigatoxin_subtypes_raw.append(item[1])
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234 # partial matches
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235 for subtype in shigatoxin_typing:
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236 for item in shigatoxin_types:
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237 if item[0] == subtype[0] and item[1] not in shigatoxin_subtypes_raw:
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238 if item[1][0:4] == "stx1":
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239 shigatoxin_subtypes.append(item[1] + "(" + str(float(subtype[1])) + ")")
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240 shigatoxin_subtypes_raw.append(item[1])
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241 if item[1][0:4] == "stx2":
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242 shigatoxin_subtypes.append(item[1] + "(" + str(float(subtype[1])) + ")")
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243 shigatoxin_subtypes_raw.append(item[1])
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244 shigatoxin_subtypes.sort()
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245 str_shigatoxin_subtype = " ".join(shigatoxin_subtypes)
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246 report.write("%s" % str_shigatoxin_subtype)
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247 report.write("</p>\n")
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248 # Quality Check
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249 disclaimer = False
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250 if any("-" in s for s in sequence_typing) or any("?" in s for s in sequence_typing):
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251 disclaimer = True
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252 if disclaimer:
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253 report.write("<p style='font-weight:bold;color:red'>Disclaimer: The data analysed do not fulfill minimum quality parameters, please consider repeating the sequencing.</p>\n")
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254 report.write("<hr/><h3>Raw data quality check</h3>\n")
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255 if args.input2:
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256 report.write("<p>FASTQC result forward: <a href='%s/datasets/%s/display/?preview=True'>Webpage</a></p>\n" % (BASE_URL, args.html1_id))
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257 report.write("<p>FASTQC result reverse: <a href='%s/datasets/%s/display/?preview=True'>Webpage</a></p>\n" % (BASE_URL, args.html2_id))
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258 else:
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259 report.write("<p>FASTQC result: <a href='%s/datasets/%s/display/?preview=True'>Webpage</a></p>\n" % (BASE_URL, args.html1_id))
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260 if args.serotyping:
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261 report.write("<br/><hr/><h3>Serotyping</h3>\n")
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262 insertFileAsTable("serogroup_OH_fcd", report, True)
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263 report.write("<br/><hr/><h3>Multi Locus Sequence Typing</h3>\n")
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264 if len(sequence_typing) > 1:
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265 insertTable(sequence_typing, report, True)
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266 if args.virulotyping:
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267 report.write("<br/><hr/><h3>Virulotyping</h3>\n")
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268 report.write("<p>This table is filtered for results with >90%% gene coverage, unfiltered results can be found <a href='%s/datasets/%s/display/?preview=True'>here</a></p>\n" % (BASE_URL, args.virulotyper_id))
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269 insertFileAsTable("pathotyper_rep_tab", report, True, "table table-cross")
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270 if args.shigatoxintyping:
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271 report.write("<br/><hr/><h3>Shiga toxin typing</h3>\n")
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272 insertFileAsTable(args.stx, report, True)
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273 if args.amrtyping:
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274 report.write("<br/><hr/><h3>AMR typing</h3>\n")
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275 report.write("<p>AMR result: <a href='%s/datasets/%s/display/?preview=True'>Webpage</a></p>\n" % (BASE_URL, args.amr_id))
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276 # write tail html
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277 insertFile(TOOL_DIR + "/report_tail.html", report)
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278 finally:
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279 report.close()
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280
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281 if __name__ == "__main__":
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282 __main__()