Dev

pathogenprofiler/__init__.py

@@ -10,4 +10,4 @@
 from .cli import *
 from .rules import *
 from .models import *
-__version__ = "4.3.0"
+__version__ = "4.4.0"

pathogenprofiler/bam.py

@@ -85,9 +85,16 @@ def run_delly(self,ref_file: str,bed_file: str) -> Vcf:
         self.bed_file = bed_file
         self.ref_file = ref_file
         if self.platform=="illumina":
-            run_cmd("delly call -t DEL -g %(ref_file)s %(bam_file)s -o %(prefix)s.delly.bcf" % vars(self))            
+            cmd = "delly call -t DEL -g %(ref_file)s %(bam_file)s -o %(prefix)s.delly.bcf" % vars(self)
+            result = run_cmd(cmd, exit_on_error=False)
         else:
-            run_cmd("delly lr -t DEL -g %(ref_file)s %(bam_file)s -o %(prefix)s.delly.bcf" % vars(self))
+            cmd = "delly lr -t DEL -g %(ref_file)s %(bam_file)s -o %(prefix)s.delly.bcf" % vars(self)
+            result = run_cmd(cmd, exit_on_error=False)
+
+        exitcode = result.returncode
+        if exitcode!=0:
+            logging.error("Delly failed, skipping")
+            return None
 
         run_cmd("bcftools view -c 2 %(prefix)s.delly.bcf | bcftools view -e '(INFO/END-POS)>=100000' -Oz -o %(prefix)s.delly.vcf.gz" % vars(self))
         run_cmd("bcftools index %(prefix)s.delly.vcf.gz" % vars(self))

pathogenprofiler/cli.py

@@ -1,7 +1,7 @@
 from .fastq import Fastq
 from .utils import run_cmd, cmd_out
 from .bam import Bam
-from .db import get_db
+from .db import get_db, check_db_exists
 from .fasta import Fasta, Paf
 from .profiler import vcf_profiler, bam_barcoder, vcf_barcoder
 import logging
@@ -335,6 +335,7 @@ def set_species(args: argparse.Namespace) -> SpeciesPrediction:
     >>> species_prediction.species
     [Species(species='Mycobacterium abscessus', prediction_info=None)]
     """
+    check_db_exists(args.software_name,args.resistance_db)
     conf = get_db(args.software_name,args.resistance_db)
     species = Species(
         species=conf["species"]

pathogenprofiler/db.py

@@ -126,7 +126,7 @@ def write_bed(db: dict,gene_dict: dict,gene_info: List[Gene],ref_file: str,outfi
         if len(drugs)==0:
             drugs = "None"
         else:
-            drugs = ",".join(drugs)
+            drugs = ",".join(sorted(list(drugs)))
         lines.append([
             gene_info[gene].chrom,
             str(genome_start),
@@ -574,12 +574,30 @@ def check_db_files(variables):
             if not os.path.isfile(val+".fai"):
                 index_ref(val)
 
+def is_db_path(string):
+    if "/" in string:
+        return True
+    elif os.path.isfile(string+".variables.json"):
+        return True
+    return False
+
+def check_db_exists(software_name,db_name):
+    db = get_db(software_name=software_name,db_name=db_name,verbose=False)
+    if db is None:
+        share_path = f"{sys.base_prefix}/share/{software_name}/"
+        logging.error(f"DB {db_name} does not exist in the current directory or in {share_path}")
+        raise FileExistsError
+
 
-def get_db(software_name,db_name):
-    if "/" in db_name:
-        share_path = "/".join(db_name.split("/")[:-1])
-        db_name = db_name.split("/")[-1]
-        variable_file_name = f"{share_path}/{db_name}.variables.json"
+def get_db(software_name,db_name,verbose=True):
+    if is_db_path(db_name):
+        if "/" in db_name:
+            share_path = "/".join(db_name.split("/")[:-1])
+            db_name = db_name.split("/")[-1]
+            variable_file_name = f"{share_path}/{db_name}.variables.json"
+        else:
+            share_path = '.'
+            variable_file_name = f"{share_path}/{db_name}.variables.json"
     else:
         share_path = f"{sys.base_prefix}/share/{software_name}/"
         variable_file_name = get_variable_file_name(software_name,db_name)
@@ -588,7 +606,8 @@ def get_db(software_name,db_name):
         return None
     variables = json.load(open(variable_file_name))
     for key,val in variables['files'].items():
-        logging.info(f"Using {key} file: {share_path}/{val}")
+        if verbose:
+            logging.info(f"Using {key} file: {share_path}/{val}")
         if ".json" in val:
             variables[key] = json.load(open(f"{share_path}/{val}"))
         elif key=='rules':
@@ -608,9 +627,13 @@ def list_db(software_name):
 
 
 def create_species_db(args,extra_files = None):
+    variables = json.load(open("variables.json"))
     if not extra_files:
         extra_files = {}
-    version = {"name":args.prefix}
+    version = {
+        "db-schema-version":variables['db-schema-version'],
+        "name":args.prefix
+    }
     if os.path.isdir('.git'):
         for l in pp.cmd_out("git log | head -4"):
             row = l.strip().split()
@@ -626,12 +649,12 @@ def create_species_db(args,extra_files = None):
         shutil.copyfile(file,target)
 
     variables_file = args.prefix+".variables.json"
-    variables = {
+    variables.update({
         "version": version,
         "files":{
             "variables": variables_file
         }
-    }
+    })
 
     if extra_files:
         for key,val in extra_files.items():

pathogenprofiler/hgvs.py

@@ -332,4 +332,30 @@ def verify_mutation_list(hgvs_mutations: List[dict], genes: List[Gene], refseq:
             converted_mutations[key] = mutation_conversion[key]
 
     logging.debug(converted_mutations)
-    return converted_mutations
+    return converted_mutations
+
+
+def split_protein_hgvs(hgvs) -> List[str]:
+    """
+    Split a protein HGVS into its components.
+
+    Parameters:
+    hgvs (str): The protein HGVS.
+
+    Returns:
+    List[str]: A list of protein HGVS components.
+
+    Example:
+    >>> split_protein_hgvs("p.MetAsnLys74IleGluThr")
+    ['p.Met74Ile', 'p.Asn74Glu', 'p.Lys74Thr']
+    """
+    # Remove the 'p.' prefix
+    hgvs = hgvs[2:]
+    components = []
+    aa = re.findall(r'[A-Z][a-z][a-z]', hgvs)
+    refs = aa[:len(aa)//2]
+    alts = aa[len(aa)//2:]
+    start_pos = int(re.search(r'\d+', hgvs).group())
+    for i,(ref,alt) in enumerate(zip(refs, alts)):
+        components.append(f'p.{ref}{start_pos+i}{alt}')
+    return components

pathogenprofiler/mutation_db.py

@@ -23,7 +23,7 @@ def add(self,data: Union[dict,list]) -> None:
             self.container.add(json.dumps(data))
 
     def to_dict_list(self) -> List[dict]:
-        return [json.loads(d) for d in self.container]
+        return [json.loads(d) for d in sorted(list(self.container))]
 
 
 

pathogenprofiler/profiler.py

@@ -48,7 +48,8 @@ def vcf_profiler(args: argparse.Namespace) -> List[Union[Variant,DrVariant,Gene,
     vcf_targets_file = "%s.targets.vcf.gz" % args.files_prefix
     if not vcf_is_indexed(args.vcf):
         run_cmd("bcftools index %s" % args.vcf)
-    # run_cmd("bcftools view -R %s %s -Oz -o %s" % (conf["bed"],args.vcf,vcf_targets_file))
+    if args.caller == 'freebayes-haplotype':
+        args.supplementary_bam = None
     annotated_variants = vcf_variant_profiler(conf, args.files_prefix, args.vcf, bam_for_phasing=args.supplementary_bam)
     return annotated_variants
 

pathogenprofiler/utils.py

@@ -20,6 +20,43 @@
 
 shared_dict = {}
 
+def get_version(tool):
+    cmds = {
+        'bcftools': 'bcftools --version',
+        'samtools': 'samtools --version',
+        'delly': 'delly --version',
+        'bwa': 'bwa',
+        'trimmomatic': 'trimmomatic -version',
+        'gatk': 'gatk -version',
+        'lofreq': 'lofreq version',
+        'bedtools': 'bedtools --version',
+        'minimap2': 'minimap2 --version',
+        'freebayes': 'freebayes --version',
+        'pilon': 'pilon --version',
+        'snpEff': 'snpEff -version',
+        'kmc': 'kmc',
+        'sourmash': 'sourmash --version',
+    }
+    regex = {
+        'bcftools': r'bcftools (\d+\.\d+\.?\d?)',
+        'samtools': r'samtools (\d+\.\d+\.?\d?)',
+        'delly': r'Delly version: v(\d+\.\d+\.?\d?)',
+        'bwa': r'Version: (\d+\.\d+\.?\d?)',
+        'trimmomatic': r'(\d+\.\d+)',
+        'gatk': r'The Genome Analysis Toolkit \(GATK\) v(\d+\.\d+\.?\d?)',
+        'lofreq': r'version: (\d+\.\d+\.?\d?)',
+        'bedtools': r'bedtools v(\d+\.\d+\.?\d?)',
+        'minimap2': r'(\d+\.\d+)',
+        'freebayes': r'version:  v(\d+\.\d+\.?\d?)',
+        'pilon': r'Pilon version (\d+\.\d+\.?\d?)',
+        'snpEff': r'SnpEff\t(\d+\.\d+.?)',
+        'kmc': r'K-Mer Counter \(KMC\) ver. (\d+\.\d+\.\d+)',
+        'sourmash': r'sourmash (\d+\.\d+\.\d?)',
+    }
+    x = sp.run([cmds[tool]], shell=True, stdout=sp.PIPE, stderr=sp.PIPE)
+    text = x.stdout.decode('utf-8') + x.stderr.decode('utf-8')
+    version = re.search(regex[tool], text).group(1)
+    return version
 
 class TempFilePrefix(object):
     """Create a temporary file prefix"""
@@ -482,10 +519,10 @@ def add_arguments_to_self(self,args: dict) -> None:
 
 
 
-def run_cmd(cmd: str, desc=None, log: str=None) -> sp.CompletedProcess:
+def run_cmd(cmd: str, desc=None, log: str=None, exit_on_error: bool=True) -> sp.CompletedProcess:
     if desc:
         logging.info(desc)
-    programs = set([x.strip().split()[0] for x in re.split("[|&;]",cmd) if x!=""])
+    programs = set([x.strip().split()[0] for x in re.split("[|&;]",cmd.strip()) if x!=""])
     missing = [p for p in programs if which(p)==False]
     if len(missing)>0:
         raise ValueError("Cant find programs: %s\n" % (", ".join(missing)))
@@ -495,7 +532,8 @@ def run_cmd(cmd: str, desc=None, log: str=None) -> sp.CompletedProcess:
     result = sp.run(cmd,shell=True,stderr=output,stdout=output)
     if result.returncode != 0:
         logging.error(result.stderr.decode("utf-8"))
-        raise ValueError("Command Failed:\n%s\nstderr:\n%s" % (cmd,result.stderr.decode()))
+        if exit_on_error:
+            raise ValueError("Command Failed:\n%s\nstderr:\n%s" % (cmd,result.stderr.decode()))
     return result
 
 def cmd_out(cmd: str) -> Generator[str, None, None]:

pathogenprofiler/variant_calling.py

@@ -130,3 +130,19 @@ def call_variants(self) -> Vcf:
             raise NotImplementedError("%s not implemented for %s platform" % (self.__software__,self.platform))
         return self.run_calling(self.calling_cmd)
 
+class FreebayesHaplotypeCaller(VariantCaller):
+    __software__ = "freebayes-haplotype"
+    def call_variants(self) -> Vcf:
+        # Call variants using Lofreq
+
+        self.calling_cmd = """
+            samtools view -T %(ref_file)s  -h %(bam_file)s {region} %(samclip_cmd)s | \
+            samtools view -b > %(temp_file_prefix)s.{region_safe}.tmp.bam && \
+            samtools index %(temp_file_prefix)s.{region_safe}.tmp.bam && \
+            freebayes -f %(ref_file)s -r {region} %(calling_params)s  %(temp_file_prefix)s.{region_safe}.tmp.bam | \
+            bcftools view -Oz -o %(temp_file_prefix)s.{region_safe}.tmp.vcf.gz && \
+            tabix %(temp_file_prefix)s.{region_safe}.tmp.vcf.gz && \
+            freebayes -f %(ref_file)s -r {region} %(calling_params)s  %(temp_file_prefix)s.{region_safe}.tmp.bam --haplotype-length 100 --haplotype-basis-alleles %(temp_file_prefix)s.{region_safe}.tmp.vcf.gz | bcftools norm -a -Oz -o %(temp_file_prefix)s.{region_safe}.vcf.gz
+            """ % vars(self)
+
+        return self.run_calling(self.calling_cmd)

pathogenprofiler/vcf.py

@@ -135,10 +135,11 @@ def run_snpeff(self,db,ref_file,gff_file,rename_chroms = None, split_indels=True
                     self.phasing_bam = f"--bam {bam_for_phasing}"
                 else:
                     self.phasing_bam = ""
-                run_cmd("bcftools view -c 1 -a %(filename)s | bcftools view -v snps | combine_vcf_variants.py --ref %(ref_file)s --gff %(gff_file)s %(phasing_bam)s | %(rename_cmd)s snpEff ann %(snpeff_data_dir_opt)s -noLog -noStats %(db)s - %(re_rename_cmd)s | bcftools sort -Oz -o %(tmp_file1)s && bcftools index %(tmp_file1)s" % vars(self))
-                run_cmd("bcftools view -c 1 -a %(filename)s | bcftools view -v indels | realign_tandem_deletions.py - %(ref_file)s %(gff_file)s - | %(rename_cmd)s snpEff ann %(snpeff_data_dir_opt)s -noLog -noStats %(db)s - %(re_rename_cmd)s | bcftools sort -Oz -o %(tmp_file2)s && bcftools index %(tmp_file2)s" % vars(self))
-                run_cmd("bcftools view -c 1 -a %(filename)s | bcftools view -v other | realign_tandem_deletions.py - %(ref_file)s %(gff_file)s - | %(rename_cmd)s snpEff ann %(snpeff_data_dir_opt)s -noLog -noStats %(db)s - %(re_rename_cmd)s | bcftools sort -Oz -o %(tmp_file3)s && bcftools index %(tmp_file3)s" % vars(self))
-                run_cmd("bcftools concat -a %(tmp_file1)s %(tmp_file2)s %(tmp_file3)s | bcftools sort -Oz -o %(vcf_csq_file)s" % vars(self))
+                run_cmd("bcftools view -c 1 -a %(filename)s | realign_tandem_deletions.py - %(ref_file)s %(gff_file)s - |  combine_vcf_variants.py --ref %(ref_file)s --gff %(gff_file)s %(phasing_bam)s | %(rename_cmd)s snpEff ann %(snpeff_data_dir_opt)s -noLog -noStats %(db)s - %(re_rename_cmd)s | bcftools sort -Oz -o %(vcf_csq_file)s" % vars(self))
+                # run_cmd("bcftools view -c 1 -a %(filename)s | bcftools view -v snps | combine_vcf_variants.py --ref %(ref_file)s --gff %(gff_file)s %(phasing_bam)s | %(rename_cmd)s snpEff ann %(snpeff_data_dir_opt)s -noLog -noStats %(db)s - %(re_rename_cmd)s | bcftools sort -Oz -o %(tmp_file1)s && bcftools index %(tmp_file1)s" % vars(self))
+                # run_cmd("bcftools view -c 1 -a %(filename)s | bcftools view -v indels | realign_tandem_deletions.py - %(ref_file)s %(gff_file)s - | %(rename_cmd)s snpEff ann %(snpeff_data_dir_opt)s -noLog -noStats %(db)s - %(re_rename_cmd)s | bcftools sort -Oz -o %(tmp_file2)s && bcftools index %(tmp_file2)s" % vars(self))
+                # run_cmd("bcftools view -c 1 -a %(filename)s | bcftools view -v other | realign_tandem_deletions.py - %(ref_file)s %(gff_file)s - | %(rename_cmd)s snpEff ann %(snpeff_data_dir_opt)s -noLog -noStats %(db)s - %(re_rename_cmd)s | bcftools sort -Oz -o %(tmp_file3)s && bcftools index %(tmp_file3)s" % vars(self))
+                # run_cmd("bcftools concat -a %(tmp_file1)s %(tmp_file2)s %(tmp_file3)s | bcftools sort -Oz -o %(vcf_csq_file)s" % vars(self))
         else :
             run_cmd("bcftools view %(filename)s | %(rename_cmd)s snpEff ann %(snpeff_data_dir_opt)s -noLog -noStats %(db)s - %(re_rename_cmd)s | bcftools view -Oz -o %(vcf_csq_file)s" % vars(self))
         return Vcf(self.vcf_csq_file,self.prefix)
@@ -186,7 +187,7 @@ def load_ann(
         variants = []
         vcf = pysam.VariantFile(self.filename)
         for var in vcf:
-            logging.debug(str(var)[:500])
+            logging.debug(str(var)[:5000])
             chrom = var.chrom
             pos = var.pos
             ref = var.ref
@@ -211,14 +212,23 @@ def load_ann(
             ann_list = [x.split("|") for x in ann_strs]
             for alt in alleles[1:]:
                 strand_support = get_stand_support(var,alt)
-
+                if strand_support[0] != None:
+                    freq = sum(strand_support)/sum(ad)
+                else:
+                    if 'QNAME' in var.info:
+                        freq = 1.0
+                    elif sv:
+                        freq = af_dict[alt]
+                    else:
+                        raise NotImplementedError
+
                 tmp_var = Variant(
                     chrom = chrom,
                     pos = int(pos),
                     ref = ref,
                     alt = alt,
                     depth = sum(ad),
-                    freq = af_dict[alt],
+                    freq = freq,
                     forward_reads = strand_support[0],
                     reverse_reads = strand_support[1],
                     sv = sv,