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4130678 +Chromosome 4135452 4135453 +Chromosome 4135456 4135457 +Chromosome 4135459 4135462 +Chromosome 4135464 4135470 +Chromosome 4139531 4139532 +Chromosome 4139534 4139535 +Chromosome 4139536 4139537 +Chromosome 4139538 4139539 +Chromosome 4143727 4143728 +Chromosome 4149159 4149160 +Chromosome 4149161 4149221 +Chromosome 4155510 4155511 +Chromosome 4155525 4155529 +Chromosome 4156802 4156911 +Chromosome 4156915 4156926 +Chromosome 4175395 4175396 +Chromosome 4175397 4175414 +Chromosome 4176383 4176392 +Chromosome 4183891 4183895 +Chromosome 4189233 4189234 +Chromosome 4189235 4189236 +Chromosome 4189237 4189272 +Chromosome 4189961 4189967 +Chromosome 4190311 4190312 +Chromosome 4190314 4190347 +Chromosome 4190348 4190376 +Chromosome 4197138 4197140 +Chromosome 4198611 4198616 +Chromosome 4201199 4201310 +Chromosome 4212832 4212836 +Chromosome 4221070 4221072 +Chromosome 4222212 4222213 +Chromosome 4222214 4222309 +Chromosome 4222322 4222323 +Chromosome 4231228 4231230 +Chromosome 4231869 4231953 +Chromosome 4253015 4253367 +Chromosome 4253876 4254214 +Chromosome 4254215 4254217 +Chromosome 4274515 4274520 +Chromosome 4298230 4298231 +Chromosome 4303498 4303499 +Chromosome 4305063 4305066 +Chromosome 4310058 4310059 +Chromosome 4310060 4310242 +Chromosome 4318481 4318482 +Chromosome 4318483 4318811 +Chromosome 4318898 4319243 +Chromosome 4320135 4320151 +Chromosome 4320385 4320386 +Chromosome 4320388 4320391 +Chromosome 4320392 4320397 +Chromosome 4320399 4320400 +Chromosome 4320401 4320402 +Chromosome 4320406 4320408 +Chromosome 4320411 4320412 +Chromosome 4322039 4322041 +Chromosome 4328212 4328214 +Chromosome 4340881 4340883 +Chromosome 4340890 4341034 +Chromosome 4350748 4350754 +Chromosome 4353151 4353922 +Chromosome 4353923 4353924 +Chromosome 4358643 4358671 +Chromosome 4358687 4358688 +Chromosome 4358867 4358868 +Chromosome 4358869 4358871 +Chromosome 4358874 4359183 +Chromosome 4361006 4361136 +Chromosome 4368536 4368662 +Chromosome 4369462 4369709 +Chromosome 4370537 4370538 +Chromosome 4372452 4372453 +Chromosome 4375614 4375646 +Chromosome 4375660 4375661 +Chromosome 4375693 4375711 +Chromosome 4379044 4379049 +Chromosome 4383144 4383145 +Chromosome 4385743 4385744 +Chromosome 4400660 4400663 +Chromosome 4411494 4411532 diff --git a/db/tbdb.rules.txt b/db/tbdb.rules.txt index ca49686..087889b 100644 --- a/db/tbdb.rules.txt +++ b/db/tbdb.rules.txt @@ -1,2 +1,2 @@ -Variant(gene_name=mmpL5,type=lof) inactivates_resistance Variant(gene_name=mmpR5) -Variant(gene_name=eis,type=lof) inactivates_resistance Variant(gene_name=eis) +Variant(gene_name=mmpL5,type=lof) inactivates_resistance Variant(gene_name=mmpR5) Loss of function variant found in mmpL5 which can abrogate the effect of genetically linked AwR and AwRI mmpR5 (Rv0678) resistance mutations. +Variant(gene_name=eis,type=lof) inactivates_resistance Variant(gene_name=eis) Loss of function variant found in eis which can abrogate the effect of genetically linked AwR and AwRI eis resistance mutations. diff --git a/tb-profiler b/tb-profiler index 85de7cd..f59dce1 100644 --- a/tb-profiler +++ b/tb-profiler @@ -21,6 +21,7 @@ from rich.logging import RichHandler import importlib import pkgutil + discovered_plugins = { name: importlib.import_module(name) for finder, name, ispkg @@ -28,10 +29,7 @@ discovered_plugins = { if name.startswith('tbprofiler_') } -docx_plugins = { - plugin.docx.__docx_template_name__:plugin for plugin in discovered_plugins.values() - if hasattr(plugin, 'docx') -} + __softwarename__ = 'tbprofiler' @@ -115,11 +113,19 @@ def main_profile(args): tbp.process_tb_profiler_args(args) variants_profile = pp.run_profiler(args) + + notes = set() if 'rules' in args.conf: - rules_applied = pp.apply_rules(args.conf['rules'], variants_profile) + rules_applied = pp.apply_rules(args.conf['rules'], variants_profile, just_make_note=True if args.implement_rules==False else False) + for var in variants_profile: + for ann in var.annotation: + if 'note' in ann: + notes.add(ann['note']) + else: rules_applied = [] + logging.debug(rules_applied) tbp.clean_up_duplicate_annotations(variants_profile) # Convert variant objects to DrVariant if they cause resistance for var in variants_profile: @@ -142,9 +148,6 @@ def main_profile(args): else: qc = pp.run_vcf_qc(args) - notes = [] - for rule in rules_applied: - notes.append(f"Rule applied: {rule}") result = tbp.create_resistance_result( args = args, @@ -192,6 +195,8 @@ def main_profile(args): "%s.bam.bai" % args.files_prefix: "%s/bam/%s.bam.bai" % (args.dir,args.prefix), "%s.nwk" % args.files_prefix: "%s/results/%s.nwk" % (args.dir,args.prefix), } + if args.save_low_dp_mask: + result_files["%s.%s.mask.bed" % (args.files_prefix,args.prefix)] = "%s/results/%s.mask.bed" % (args.dir,args.prefix) for file,target in result_files.items(): if os.path.isfile(file): shutil.move(file,target) @@ -350,6 +355,7 @@ def int_2_or_more(arg): variant_callers = [cls.__software__ for cls in vc.VariantCaller.__subclasses__()] +docx_plugins = [cls.__template_name__ for cls in tbp.docx.DocxResultTemplate.__subclasses__()] @@ -380,7 +386,7 @@ output.add_argument('--txt',action="store_true",help="Add text output") output.add_argument('--text_template','--text-template',type=str,help='Jinja2 formatted template for output') output.add_argument('--docx',action="store_true",help="Add docx output") output.add_argument('--docx_template','--docx-template',help="Supply custom template for --docx output") -output.add_argument('--docx_plugin','--docx-plugin',choices=list(docx_plugins),help="Use a plugin template for --docx output") +output.add_argument('--docx_plugin','--docx-plugin',choices=docx_plugins,help="Use a plugin template for --docx output") output.add_argument('--add_columns','--add-columns',default=None,type=str,help="Add additional columns found in the mutation database to the text and csv results") output.add_argument('--add_mutation_metadata','--add-mutation-metadata',action="store_true",help=argparse.SUPPRESS) output.add_argument('--dir','-d',default=".",help='Storage directory') @@ -420,12 +426,15 @@ algorithm.add_argument('--no_lineage','--no-lineage',action="store_true",help=ar algorithm.add_argument('--add_variant_annotations','--add-variant-annotations',action="store_true",help=argparse.SUPPRESS) algorithm.add_argument('--threads','-t',default=1,help='Threads to use',type=int) algorithm.add_argument('--ram',default=2,help='Maximum memory to use',type=int) +algorithm.add_argument('--implement_rules','--implement-rules',action="store_true",help='Use rules implemented in the resistance library (by default only a note will be made)') other=parser_sub.add_argument_group("Other options") other.add_argument('--logging',type=str.upper,default="INFO",choices=["DEBUG","INFO","WARNING","ERROR","CRITICAL"],help='Logging level') other.add_argument('--debug',action='store_true',help=argparse.SUPPRESS) other.add_argument('--delly_vcf','--delly-vcf',help=argparse.SUPPRESS) other.add_argument('--supplementary_bam','--supplementary-bam',help=argparse.SUPPRESS) +other.add_argument('--low_dp_mask','--low-dp-mask',help=argparse.SUPPRESS) +other.add_argument('--save_low_dp_mask','--save-low-dp-mask',action='store_true',help=argparse.SUPPRESS) other.add_argument('--no_clean','--no-clean', action='store_true',help=argparse.SUPPRESS) other.add_argument('--temp',help="Temp firectory to process all files",type=str,default=".") other.add_argument('--version', action='version', version="tb-profiler version %s" % tbp.__version__) diff --git a/tbprofiler/__init__.py b/tbprofiler/__init__.py index 5441640..f966fdd 100644 --- a/tbprofiler/__init__.py +++ b/tbprofiler/__init__.py @@ -6,5 +6,6 @@ from .output import * from .snp_dists import * from .phylo import * +from .docx import * -__version__ = "6.3.0" +__version__ = "6.4.0" diff --git a/tbprofiler/docx.py b/tbprofiler/docx.py index fb4dc5e..2911d27 100644 --- a/tbprofiler/docx.py +++ b/tbprofiler/docx.py @@ -1,11 +1,22 @@ import sys from docxtpl import DocxTemplate from collections import defaultdict -from .models import ProfileResult +from .models import ProfileResult, VcfQC from docx import Document from typing import List from copy import deepcopy import logging +from abc import abstractmethod +from .utils import rv2genes, get_gene2drugs +import os + + + +class DocxResultTemplate: + @abstractmethod + def write_output(self, result: ProfileResult, conf: dict, outfile: str): + """Write the output to the Word document""" + pass def sanitize(d): d = d.replace("-","_") @@ -33,11 +44,16 @@ def _merge_cells(tab, rows: List[int], column: int): return c1 = tab.cell(rows[0], column) c2 = tab.cell(rows[-1], column) - + c1_font_size = c1.paragraphs[0].runs[0].font.size + for r in rows[1:]: - tab.cell(r, column).text = "" + tab.cell(r, column).text = '' + + cm = c1.merge(c2) + + values_in_next_column = set([tab.rows[r].cells[column+1].text for r in rows]) for val in values_in_next_column: rows_with_val = [r for r in rows if tab.c[r][column+1].text == val] @@ -54,90 +70,261 @@ def _merge_cells(tab, rows: List[int], column: int): doc.save(filename) +class DefaultTemplate(DocxResultTemplate): + __template_name__ = "default" + def __init__(self, *args, **kwargs): + super().__init__(*args, **kwargs) + __dir__ = os.path.dirname(os.path.abspath(__file__)) + # self.template_filename = f"{__dir__}/template.docx" + self.template_filename = sys.prefix+"/share/tbprofiler/default_template.docx" + def write_output( + self, + result: ProfileResult, + conf: dict, + output_filename: str): + + id2name = rv2genes(conf['bed']) + db = conf['json_db'] + tier1_genes = set() + for gene in db: + for var in db[gene]: + for ann in db[gene][var]['annotations']: + if ann['type']=='drug_resistance': + tier1_genes.add(id2name[gene]) + + tier1_genes.add('mmpL5') + + if isinstance(result.qc, VcfQC): + qc_check = {d:100 for d in id2name.values()} + result.notes.append("This report was generated from a VCF where gene coverage is not available. It is assumed all genes are covered well sufficiently.") + else: + qc_check = {d.target:d.percent_depth_pass for d in result.qc.target_qc} + + + gene2drugs = get_gene2drugs(conf['bed']) + table = [{'name':d, 'genes':[]} for d in conf['drugs']] + for gene,drugs in gene2drugs.items(): + if gene not in tier1_genes: continue + for d in drugs: + item = [i for i in table if i['name']==d][0] + item['genes'].append({'name':gene, 'variants':[]}) + + for var in result.dr_variants: + for d in var.drugs: + item = [i for i in table if i['name']==d['drug']][0] + gene_item = [i for i in item['genes'] if i['name']==var.gene_name][0] + gene_item['variants'].append({ + 'change':var.change, + 'freq':var.freq, + 'confidence':d['confidence'], + 'comment':d['comment'] + }) + + comments = {} + rows = [] + for d in table: + for g in d['genes']: + if len(g['variants']) == 0: + rows.append({ + 'drug':d['name'].replace("_"," ").title(), + 'gene':g['name'], + 'qc':qc_check[g['name']], + 'change':'', + 'freq':'', + 'confidence':'', + 'comment':'' + }) + else: + for v in g['variants']: + if v['comment']!="" and v['comment'] not in comments: + comments[v['comment']] = len(comments) + 1 + rows.append({ + 'drug':d['name'].replace("_"," ").title(), + 'gene':g['name'], + 'qc':qc_check[g['name']], + 'change':v['change'], + 'freq':int(v['freq']*100), + 'confidence':v['confidence'] if v['confidence']!="" else "Uncertain significance", + 'comment':comments.get(v['comment'],'') + }) + + resistant_drugs_tmp = [r['drug'] for r in rows if r['change']!=""] + resistant_drugs = [d for d in conf['drugs'] if d.title() in resistant_drugs_tmp] + if len(resistant_drugs)>0: + result_summary = f"Known resistance variants for {', '.join(resistant_drugs[:-1])} and {resistant_drugs[-1]} detected." + else: + result_summary = "No known resistance variants detected." + + + poor_coverage_genes = [g for g in qc_check if qc_check[g]<99] + if len(poor_coverage_genes)>0: + result.notes.append(f"Insufficient coverage detected in {len(poor_coverage_genes)} genes.") + + fail_variants = [] + fail_comments = {} + for var in result.qc_fail_variants: + if not hasattr(var,'drugs'): continue + for d in var.drugs: + + if d['comment']!="" and d['comment'] not in fail_comments: + fail_comments[d['comment']] = len(fail_comments) + 1 + fail_variants.append({ + 'drug':d['drug'], + 'gene':var.gene_name, + 'qc':qc_check[var.gene_name], + 'change':var.change, + 'confidence':d['confidence'], + 'freq':int(var.freq*100), + 'depth':var.depth, + 'comment':fail_comments.get(d['comment'],'') + }) + if var.gene_name in ('mmpL5','mmpS5') and var.type=='frameshift_variant': + for ann in var.annotation: + if ann['type']=='who_confidence' and ann['original_mutation']=='LoF': + fail_variants.append({ + 'drug':ann['drug'], + 'gene':var.gene_name, + 'qc':qc_check[var.gene_name], + 'change':var.change, + 'confidence':ann['confidence'], + 'freq':int(var.freq*100), + 'depth':var.depth, + 'comment':fail_comments.get(ann['comment'],'') + }) + + fail_variants_unique = set([(f['gene'],f['change']) for f in fail_variants]) + if len(fail_variants)>0: + result.notes.append(f"{len(fail_variants_unique)} resistance variants failed QC checks. These have not been used to generate the mutation report. See QC failed variants table for details.") + + + other_variants = [] + other_comments = {} + for var in result.other_variants: + drug_annotation = {a['drug']:a for a in var.annotation if a['type']=='who_confidence'} + for drug_name in var.gene_associated_drugs: + d = drug_annotation.get(drug_name,{}) + if d.get('comment','')!='' and d['comment'] not in other_comments: + other_comments[d['comment']] = len(other_comments) + 1 + other_variants.append({ + 'drug':drug_name, + 'gene':var.gene_name, + 'qc':qc_check[var.gene_name], + 'change':var.change, + 'confidence':d.get('confidence','Uncertain significance'), + 'freq':int(var.freq*100), + 'depth':var.depth, + 'comment':other_comments.get(d.get('comment',''),'') + }) + + + + context = { + 'd': result.model_dump(), + 'result_summary': result_summary, + 'rows':rows, + 'comments': comments, + 'qc_check': qc_check, + 'notes': ' '.join(result.notes), + 'fail_variants': fail_variants, + 'fail_comments': fail_comments, + 'other_variants': other_variants, + 'other_comments': other_comments + } + + tpl = DocxTemplate(self.template_filename) + tpl.render(context) + tpl.save(output_filename) + + merge_cells(output_filename) + + def write_docx(result: ProfileResult,conf,outfile,template_file = None, plugin = None): if template_file is None: template_file = sys.prefix+"/share/tbprofiler/default_template.docx" if plugin: - variables = plugin.docx.create_output(result, conf) + plugin_cls = plugin() + plugin_cls.write_output(result, conf,outfile) else: - dr_variant_table = [] - comments = [] + output_cls = DefaultTemplate() + output_cls.write_output(result, conf, outfile) + # dr_variant_table = [] + # comments = [] - for var in result.dr_variants: - for drug in var.drugs: - mutation = var.change if len(var.change) < 15 else var.change[:11]+"..."+var.change[-3:] - if drug['comment'] not in comments and len(drug['comment']): - comments.append(drug['comment']) - dr_variant_table.append({ - 'drug': drug['drug'], - 'gene': var.gene_name, - 'mutation': mutation, - 'depth': var.depth, - 'frequency': round(var.freq * 100,2), - 'confidence': drug['confidence'], - 'comment': comments.index(drug['comment'])+1 if len(drug['comment']) else "" - }) - drugs_found = set([x['drug'] for x in dr_variant_table]) - for drug in conf['drugs']: - if drug not in drugs_found: - dr_variant_table.append({ - 'drug': drug, - 'gene': "", - 'mutation': "", - 'depth': "", - 'frequency': "", - 'confidence': "", - 'comment': "" - }) + # for var in result.dr_variants: + # for drug in var.drugs: + # mutation = var.change if len(var.change) < 15 else var.change[:11]+"..."+var.change[-3:] + # if drug['comment'] not in comments and len(drug['comment']): + # comments.append(drug['comment']) + # dr_variant_table.append({ + # 'drug': drug['drug'], + # 'gene': var.gene_name, + # 'mutation': mutation, + # 'depth': var.depth, + # 'frequency': round(var.freq * 100,2), + # 'confidence': drug['confidence'], + # 'comment': comments.index(drug['comment'])+1 if len(drug['comment']) else "" + # }) + # drugs_found = set([x['drug'] for x in dr_variant_table]) + # for drug in conf['drugs']: + # if drug not in drugs_found: + # dr_variant_table.append({ + # 'drug': drug, + # 'gene': "", + # 'mutation': "", + # 'depth': "", + # 'frequency': "", + # 'confidence': "", + # 'comment': "" + # }) - gene_qc = [] - if 'target_qc' in result.qc: - for item in result.qc.target_qc: + # gene_qc = [] + # if 'target_qc' in result.qc: + # for item in result.qc.target_qc: - gene_qc.append({ - 'status': 'ok' if item.percent_depth_pass>0.9 else 'fail', - 'gene': item.target, - 'median_depth': item.median_depth, - 'percent_depth_pass': item.percent_depth_pass, - }) + # gene_qc.append({ + # 'status': 'ok' if item.percent_depth_pass>0.9 else 'fail', + # 'gene': item.target, + # 'median_depth': item.median_depth, + # 'percent_depth_pass': item.percent_depth_pass, + # }) - dr_variant_table = sorted(dr_variant_table,key=lambda x: conf['drugs'].index(x['drug'])) + # dr_variant_table = sorted(dr_variant_table,key=lambda x: conf['drugs'].index(x['drug'])) - other_variants_table = [] - for var in result.other_variants: - for ann in var.annotation: - other_variants_table.append({ - 'gene': var.gene_name, - 'mutation': var.change, - 'depth': var.depth, - 'frequency': round(var.freq * 100,2), - 'drug': ann['drug'], - 'confidence': ann['confidence'], - }) + # other_variants_table = [] + # for var in result.other_variants: + # for ann in var.annotation: + # other_variants_table.append({ + # 'gene': var.gene_name, + # 'mutation': var.change, + # 'depth': var.depth, + # 'frequency': round(var.freq * 100,2), + # 'drug': ann['drug'], + # 'confidence': ann['confidence'], + # }) - data = result.model_dump() + # data = result.model_dump() - variables = { - 'date':data['timestamp'].strftime("%d %b %Y"), - 'sublineage': data['sub_lineage'], - 'version': data['pipeline']['software_version'], - 'drtype': data['drtype'], - 'd': data, - 'dr_variants_table': dr_variant_table, - 'other_variants_table': other_variants_table, - 'comments': comments, - 'gene_qc': gene_qc - } + # variables = { + # 'date':data['timestamp'].strftime("%d %b %Y"), + # 'sublineage': data['sub_lineage'], + # 'version': data['pipeline']['software_version'], + # 'drtype': data['drtype'], + # 'd': data, + # 'dr_variants_table': dr_variant_table, + # 'other_variants_table': other_variants_table, + # 'comments': comments, + # 'gene_qc': gene_qc + # } - doc = DocxTemplate(template_file) - doc.render(variables) - doc.save(outfile) + # doc = DocxTemplate(template_file) + # doc.render(variables) + # doc.save(outfile) - merge_cells(outfile) \ No newline at end of file + # merge_cells(outfile) \ No newline at end of file diff --git a/tbprofiler/output.py b/tbprofiler/output.py index 317df59..6e880b8 100644 --- a/tbprofiler/output.py +++ b/tbprofiler/output.py @@ -1,7 +1,8 @@ from .text import write_text -from .docx import write_docx +from .docx import write_docx, DocxResultTemplate import logging from .models import ProfileResult +from pathogenprofiler import FastaQC, BamQC, VcfQC def write_outputs(args,result: ProfileResult,template_file: str = None): logging.info("Writing outputs") @@ -23,5 +24,7 @@ def write_outputs(args,result: ProfileResult,template_file: str = None): if args.docx: logging.info(f"Writing docx file: {docx_output}") if args.docx_plugin: - args.docx_plugin = args.plugins[args.docx_plugin] - write_docx(result,args.conf,docx_output,template_file = args.docx_template, plugin = args.docx_plugin) \ No newline at end of file + available_plugins = {cls.__template_name__:cls for cls in DocxResultTemplate.__subclasses__()} + args.docx_plugin = available_plugins[args.docx_plugin] + + write_docx(result,args.conf,docx_output,template_file = args.docx_template, plugin = args.docx_plugin) diff --git a/tbprofiler/phylo.py b/tbprofiler/phylo.py index 7fe855f..ace02d7 100644 --- a/tbprofiler/phylo.py +++ b/tbprofiler/phylo.py @@ -83,7 +83,7 @@ def prepare_usher(treefile: str,vcf_file: str) -> None: def prepare_sample_consensus(sample: str,input_vcf: str,args: argparse.Namespace) -> str: s = sample tmp_vcf = f"{args.files_prefix}.{s}.vcf.gz" - run_cmd(f"bcftools norm -m - {input_vcf} | bcftools view -T ^{args.conf['bedmask']} | bcftools filter --SnpGap 50 | bcftools view -v snps | annotate_maaf.py | bcftools filter -S . -e 'MAAF<0.7' |bcftools filter -S . -e 'FMT/DP<{args.conf['variant_filters']['depth_soft']}' | rename_vcf_sample.py --sample-name {s} | bcftools view -v snps -Oz -o {tmp_vcf}") + run_cmd(f"bcftools norm -m - {input_vcf} | bcftools view -T ^{args.conf['bedmask']} | bcftools view -v snps | annotate_maaf.py | bcftools filter -S . -e 'MAAF<0.7' |bcftools filter -S . -e 'FMT/DP<{args.conf['variant_filters']['depth_soft']}' | bcftools filter --SnpGap 50 |rename_vcf_sample.py --sample-name {s} | bcftools view -v snps -Oz -o {tmp_vcf}") run_cmd(f"bcftools index {tmp_vcf}") mask_bed = f"{args.files_prefix}.{s}.mask.bed" @@ -92,6 +92,9 @@ def prepare_sample_consensus(sample: str,input_vcf: str,args: argparse.Namespace if args.bam: generate_low_dp_mask(f"{args.bam}",args.conf['ref'],mask_bed) mask_cmd = f"-m {mask_bed} -M N" + elif args.low_dp_mask: + mask_bed = args.low_dp_mask + mask_cmd = f"-m {mask_bed} -M N" elif args.vcf: generate_low_dp_mask_vcf(args.vcf,mask_bed) mask_cmd = f"-m {mask_bed} -M N" diff --git a/tbprofiler/reformat.py b/tbprofiler/reformat.py index 02da7ba..9d78422 100644 --- a/tbprofiler/reformat.py +++ b/tbprofiler/reformat.py @@ -270,7 +270,7 @@ def clean_up_duplicate_annotations(variants: Variant) -> None: """ confidence_levels = ['Assoc w R','Assoc w R - Interim','Uncertain significance','Not assoc w R - Interim','Not assoc w R'] for var in variants: - keys = set([(ann['type'],ann['drug']) for ann in var.annotation]) + keys = sorted(list(set([(ann['type'],ann['drug']) for ann in var.annotation]))) new_annotations = [] for key in keys: confidence_anns = []