def _debug_dummy_plot(
taxonomy: Taxonomy,
htmlfile: Filename,
scoring: Scoring = Scoring.SHEL,
):
"""
Generate dummy Krona plot via Krona 2.0 XML spec and exit
"""
print(gray(f'Generating dummy Krona plot {htmlfile}...'), end='')
sys.stdout.flush()
samples: List[Sample] = [
Sample('SINGLE'),
]
krona: KronaTree = KronaTree(
samples,
min_score=Score(35),
max_score=Score(100),
scoring=scoring,
)
polytree: MultiTree = MultiTree(samples=samples)
polytree.grow(ontology=taxonomy)
polytree.toxml(ontology=taxonomy, krona=krona)
krona.tohtml(htmlfile, pretty=True)
print(green('OK!'))
def generate_krona():
"""Generate Krona plot with all the results via Krona 2.0 XML spec"""
print(gray('\nBuilding the taxonomy multiple tree... '), end='')
sys.stdout.flush()
krona: KronaTree = KronaTree(
samples,
num_raw_samples=len(raw_samples),
stats=stats,
min_score=Score(
min([
min(scores[sample].values()) for sample in samples
if len(scores[sample])
])),
max_score=Score(
max([
max(scores[sample].values()) for sample in samples
if len(scores[sample])
])),
scoring=scoring,
)
polytree.grow(ontology=ncbi,
abundances=counts,
accs=accs,
scores=scores)
print(green('OK!'))
print(gray('Generating final plot (') + magenta(htmlfile) +
gray(')... '),
end='')
sys.stdout.flush()
polytree.toxml(ontology=ncbi, krona=krona)
krona.tohtml(htmlfile, pretty=False)
print(green('OK!'))
def swmean(cnt1: int, sco1: Score, cnt2: int,
sco2: Score) -> Score:
"""Weighted mean of scores by counts"""
if sco1 == NO_SCORE:
return sco2
elif sco2 == NO_SCORE:
return sco1
return Score((cnt1 * sco1 + cnt2 * sco2) / (cnt1 + cnt2))
def read_output(
output_file: Filename,
scoring: Scoring = Scoring.SHEL,
minscore: Score = None,
) -> Tuple[str, SampleStats, Counter[Id], Dict[Id, Score]]:
"""
Read Centrifuge output file
Args:
output_file: output file name
scoring: type of scoring to be applied (see Scoring class)
minscore: minimum confidence level for the classification
Returns:
log string, statistics, abundances counter, scores dict
"""
output: io.StringIO = io.StringIO(newline='')
all_scores: Dict[Id, List[Score]] = {}
all_length: Dict[Id, List[int]] = {}
taxids: Set[Id] = set()
num_read: int = 0
nt_read: int = 0
num_uncl: int = 0
last_error_read: int = -1 # Number of read of the last error
num_errors: int = 0 # Number or reads discarded due to error
output.write(gray(f'Loading output file {output_file}... '))
try:
with open(output_file, 'r') as file:
file.readline() # discard header
for output_line in file:
try:
_, _, _tid, _score, _, _, _length, *_ = output_line.split(
'\t')
except ValueError:
print(
yellow('Failure'), 'parsing line elements:'
f' {output_line} in {output_file}'
'. Ignoring line!')
last_error_read = num_read + 1
num_errors += 1
continue
tid = Id(_tid)
try:
# From Centrifuge score get "single hit equivalent length"
shel = Score(float(_score)**0.5 + 15)
length = int(_length)
except ValueError:
print(yellow('Failure'), f'parsing score ({_score}) for ',
f'query length {_length} for taxid {_tid}',
f'in {output_file}. Ignoring line!')
last_error_read = num_read + 1
num_errors += 1
continue
num_read += 1
nt_read += length
if tid == UNCLASSIFIED: # Just count unclassified reads
num_uncl += 1
continue
else:
taxids.add(tid) # Save all the tids of classified reads
if minscore is not None and shel < minscore:
continue # Ignore read if low confidence
try:
all_scores[tid].append(shel)
except KeyError:
all_scores[tid] = [
shel,
]
try:
all_length[tid].append(length)
except KeyError:
all_length[tid] = [
length,
]
except FileNotFoundError:
raise Exception(red('\nERROR! ') + f'Cannot read "{output_file}"')
if last_error_read == num_read + 1: # Check error in last line: truncated!
print(yellow('Warning!'), f'{output_file} seems truncated!')
counts: Counter[Id] = col.Counter(
{tid: len(all_scores[tid])
for tid in all_scores})
output.write(green('OK!\n'))
if num_read == 0:
raise Exception(
red('\nERROR! ') +
f'Cannot read any sequence from "{output_file}"')
filt_seqs: int = sum([len(scores) for scores in all_scores.values()])
if filt_seqs == 0:
raise Exception(red('\nERROR! ') + 'No sequence passed the filter!')
# Get statistics
stat: SampleStats = SampleStats(minscore=minscore,
nt_read=nt_read,
scores=all_scores,
lens=all_length,
seq_read=num_read,
seq_unclas=num_uncl,
seq_filt=filt_seqs,
tid_clas=len(taxids))
# Output statistics
if num_errors:
output.write(
gray(' Seqs fail: ') + red(f'{num_errors:_d}\t') +
gray('(Last error in read ') + red(f'{last_error_read}') +
gray(')\n'))
output.write(
gray(' Seqs read: ') + f'{stat.seq.read:_d}\t' + gray('[') +
f'{stat.nt_read}' + gray(']\n'))
output.write(
gray(' Seqs clas: ') + f'{stat.seq.clas:_d}\t' + gray('(') +
f'{stat.get_unclas_ratio():.2%}' + gray(' unclassified)\n'))
output.write(
gray(' Seqs pass: '******'{stat.seq.filt:_d}\t' + gray('(') +
f'{stat.get_reject_ratio():.2%}' + gray(' rejected)\n'))
output.write(
gray(' Scores: min = ') + f'{stat.sco.mini:.1f}' + gray(', max = ') +
f'{stat.sco.maxi:.1f}' + gray(', avr = ') + f'{stat.sco.mean:.1f}\n')
output.write(
gray(' Length: min = ') + f'{stat.len.mini}' + gray(', max = ') +
f'{stat.len.maxi}' + gray(', avr = ') + f'{stat.len.mean}\n')
output.write(
gray(' TaxIds: by classifier = ') + f'{stat.tid.clas}' +
gray(', by filter = ') + f'{stat.tid.filt}\n')
# Select score output
out_scores: Dict[Id, Score]
if scoring is Scoring.SHEL:
out_scores = {tid: Score(mean(all_scores[tid])) for tid in all_scores}
elif scoring is Scoring.LENGTH:
out_scores = {tid: Score(mean(all_length[tid])) for tid in all_length}
elif scoring is Scoring.LOGLENGTH:
out_scores = {
tid: Score(log10(mean(all_length[tid])))
for tid in all_length
}
elif scoring is Scoring.NORMA:
scores: Dict[Id, Score] = {
tid: Score(mean(all_scores[tid]))
for tid in all_scores
}
lengths: Dict[Id, Score] = {
tid: Score(mean(all_length[tid]))
for tid in all_length
}
out_scores = {
tid: Score(scores[tid] / lengths[tid] * 100)
for tid in scores
}
else:
print(red('ERROR!'), f' Centrifuge: Unsupported Scoring "{scoring}"')
raise Exception('Unsupported scoring')
# Return
return output.getvalue(), stat, counts, out_scores
def read_clark_output(
output_file: Filename,
scoring: Scoring = Scoring.CLARK_C,
minscore: Score = None,
) -> Tuple[str, SampleStats, Counter[Id], Dict[Id, Score]]:
"""
Read CLARK(-l)(-S) full mode output file
Args:
output_file: output file name
scoring: type of scoring to be applied (see Scoring class)
minscore: minimum confidence level for the classification
Returns:
log string, statistics, abundances counter, scores dict
"""
output: io.StringIO = io.StringIO(newline='')
all_scores: Dict[Id, List[Score]] = {}
all_confs: Dict[Id, List[Score]] = {}
all_gammas: Dict[Id, List[Score]] = {}
all_length: Dict[Id, List[int]] = {}
taxids: Set[Id] = set()
num_read: int = 0
nt_read: int = 0
num_uncl: int = 0
last_error_read: int = -1 # Number of read of the last error
num_errors: int = 0 # Number or reads discarded due to error
output.write(gray(f'Loading output file {output_file}... '))
try:
with open(output_file, 'r') as file:
# Check number of cols in header
header = file.readline().split(',')
if len(header) != 8:
print(
red('\nERROR! ') + 'CLARK output format of ',
yellow(f'"{output_file}"'), 'not supported.')
print(magenta('Expected:'),
'ID,Length,Gamma,1st,score1,2nd,score2,conf')
print(magenta('Found:'), ','.join(header), end='')
print(blue('HINT:'), 'Use CLARK, CLARK-l, or CLARK-S '
'with full mode (', blue('-m 0'), ')')
raise Exception('Unsupported file format. Aborting.')
for raw_line in file:
try:
output_line = raw_line.strip()
(_label, _length, _gamma, _tid1, _score1, _tid2, _score2,
_conf) = output_line.split(',')
except ValueError:
print(
yellow('Failure'), 'parsing line elements:'
f' {output_line} in {output_file}'
'. Ignoring line!')
last_error_read = num_read + 1
num_errors += 1
continue
try:
length: int = int(_length)
gamma: Score = Score(float(_gamma))
tid1: Id = Id(_tid1)
score1: Score = Score(float(_score1))
tid2: Id = Id(_tid2)
score2: Score = Score(float(_score2))
conf: Score = Score(float(_conf))
except ValueError:
print(
yellow('Failure'), 'parsing line elements:'
f' {output_line} in {output_file}'
'. Ignoring line!')
last_error_read = num_read + 1
num_errors += 1
continue
num_read += 1
nt_read += length
# Select tid and score between CLARK assignments 1 and 2
tid: Id = tid1
score: Score = score1
if tid1 == UNCLASSIFIED:
if tid2 == UNCLASSIFIED: # Just count unclassified reads
num_uncl += 1
continue
else: # Majority of read unclassified
tid = tid2
score = score2
conf = Score(1 - conf) # Get CLARK's h2/(h1+h2)
# From CLARK_C(S) score get "single hit equivalent length"
shel: Score = Score(score + K_MER_SIZE)
taxids.add(tid) # Save all the selected tids (tid1 or tid2)
if minscore is not None: # Decide if ignore read if low score
if scoring is Scoring.CLARK_C:
if conf < minscore:
continue
elif scoring is Scoring.CLARK_G:
if gamma < minscore:
continue
else:
if shel < minscore:
continue
try:
all_scores[tid].append(shel)
except KeyError:
all_scores[tid] = [
shel,
]
try:
all_confs[tid].append(conf)
except KeyError:
all_confs[tid] = [
conf,
]
try:
all_gammas[tid].append(gamma)
except KeyError:
all_gammas[tid] = [
gamma,
]
try:
all_length[tid].append(length)
except KeyError:
all_length[tid] = [
length,
]
except FileNotFoundError:
raise Exception(red('\nERROR! ') + f'Cannot read "{output_file}"')
if last_error_read == num_read + 1: # Check error in last line: truncated!
print(yellow('Warning!'), f'{output_file} seems truncated!')
counts: Counter[Id] = col.Counter(
{tid: len(all_scores[tid])
for tid in all_scores})
output.write(green('OK!\n'))
if num_read == 0:
raise Exception(
red('\nERROR! ') +
f'Cannot read any sequence from "{output_file}"')
filt_seqs: int = sum([len(scores) for scores in all_scores.values()])
if filt_seqs == 0:
raise Exception(red('\nERROR! ') + 'No sequence passed the filter!')
# Get statistics
stat: SampleStats = SampleStats(minscore=minscore,
nt_read=nt_read,
lens=all_length,
scores=all_scores,
scores2=all_confs,
scores3=all_gammas,
seq_read=num_read,
seq_unclas=num_uncl,
seq_filt=filt_seqs,
tid_clas=len(taxids))
# Output statistics
if num_errors:
output.write(
gray(' Seqs fail: ') + red(f'{num_errors:_d}\t') +
gray('(Last error in read ') + red(f'{last_error_read}') +
gray(')\n'))
output.write(
gray(' Seqs read: ') + f'{stat.seq.read:_d}\t' + gray('[') +
f'{stat.nt_read}' + gray(']\n'))
output.write(
gray(' Seqs clas: ') + f'{stat.seq.clas:_d}\t' + gray('(') +
f'{stat.get_unclas_ratio():.2%}' + gray(' unclassified)\n'))
output.write(
gray(' Seqs pass: '******'{stat.seq.filt:_d}\t' + gray('(') +
f'{stat.get_reject_ratio():.2%}' + gray(' rejected)\n'))
output.write(
gray(' Hit (score): min = ') + f'{stat.sco.mini:.1f},' +
gray(' max = ') + f'{stat.sco.maxi:.1f},' + gray(' avr = ') +
f'{stat.sco.mean:.1f}\n')
output.write(
gray(' Conf. score: min = ') + f'{stat.sco2.mini:.1f},' +
gray(' max = ') + f'{stat.sco2.maxi:.1f},' + gray(' avr = ') +
f'{stat.sco2.mean:.1f}\n')
output.write(
gray(' Gamma score: min = ') + f'{stat.sco3.mini:.1f},' +
gray(' max = ') + f'{stat.sco3.maxi:.1f},' + gray(' avr = ') +
f'{stat.sco3.mean:.1f}\n')
output.write(
gray(' Read length: min = ') + f'{stat.len.mini},' + gray(' max = ') +
f'{stat.len.maxi},' + gray(' avr = ') + f'{stat.len.mean}\n')
output.write(
gray(' TaxIds: by classifier = ') + f'{stat.tid.clas}' +
gray(', by filter = ') + f'{stat.tid.filt}\n')
# Select score output
out_scores: Dict[Id, Score]
if scoring is Scoring.SHEL:
out_scores = {tid: Score(mean(all_scores[tid])) for tid in all_scores}
elif scoring is Scoring.CLARK_C:
out_scores = {
tid: Score(mean(all_confs[tid]) * 100)
for tid in all_confs
}
elif scoring is Scoring.CLARK_G:
out_scores = {tid: Score(mean(all_gammas[tid])) for tid in all_gammas}
elif scoring is Scoring.LENGTH:
out_scores = {tid: Score(mean(all_length[tid])) for tid in all_length}
elif scoring is Scoring.LOGLENGTH:
out_scores = {
tid: Score(log10(mean(all_length[tid])))
for tid in all_length
}
elif scoring is Scoring.NORMA:
scores: Dict[Id, Score] = {
tid: Score(mean(all_scores[tid]))
for tid in all_scores
}
lengths: Dict[Id, Score] = {
tid: Score(mean(all_length[tid]))
for tid in all_length
}
out_scores = {
tid: Score(scores[tid] / lengths[tid] * 100)
for tid in scores
}
else:
print(red('ERROR!'), f'clark: Unsupported Scoring "{scoring}"')
raise Exception('Unsupported scoring')
# Return
return output.getvalue(), stat, counts, out_scores
def read_lmat_output(
output_file: Filename,
scoring: Scoring = Scoring.LMAT,
minscore: Score = None,
) -> Tuple[str, SampleStats, Counter[Id], Dict[Id, Score]]:
"""
Read LMAT output (iterate over all the output files)
Args:
output_file: output file name (prefix)
scoring: type of scoring to be applied (see Scoring class)
minscore: minimum confidence level for the classification
Returns:
log string, abundances counter, scores dict
"""
output: io.StringIO = io.StringIO(newline='')
all_scores: Dict[Id, List[Score]] = {}
all_length: Dict[Id, List[int]] = {}
nt_read: int = 0
matchings: Counter[Match] = Counter()
output_files: List[Filename] = []
# Select files to process depending on if the output files are explicitly
# given or directory name is provided (all the output files there)
if os.path.isdir(output_file): # Just the directory name is provided
dirname = os.path.normpath(output_file)
for file in os.listdir(dirname): # Add all LMAT output files in dir
if ('_output' in file and file.endswith('.out')
and 'canVfin' not in file and 'pyLCA' not in file):
output_files.append(Filename(file))
else: # Explicit path and file name prefix is given
dirname, basename = os.path.split(output_file)
for file in os.listdir(dirname): # Add selected output files in dir
if (file.startswith(basename) and file.endswith('.out')
and 'canVfin' not in file and 'pyLCA' not in file):
output_files.append(Filename(file))
if not output_files:
raise Exception(
f'\n\033[91mERROR!\033[0m Cannot read from "{output_file}"')
# Read LMAT output files
for output_name in output_files:
path: Filename = Filename(os.path.join(dirname, output_name))
output.write(f'\033[90mLoading output file {path}...\033[0m')
try:
with open(path, 'r') as io_file:
for seq in SeqIO.parse(io_file, "lmat"):
tid: Id = seq.annotations['final_taxid']
score: Score = seq.annotations['final_score']
match: Match = Match.lmat(seq.annotations['final_match'])
matchings[match] += 1
length: int = len(seq)
nt_read += length
if minscore is not None:
if score < minscore: # Ignore read if low score
continue
if match in [Match.DIRECTMATCH, Match.MULTIMATCH]:
try:
all_scores[tid].append(score)
except KeyError:
all_scores[tid] = [
score,
]
try:
all_length[tid].append(length)
except KeyError:
all_length[tid] = [
length,
]
except FileNotFoundError:
raise Exception(red('\nERROR!') + f'Cannot read "{path}"')
output.write(green('OK!\n'))
abundances: Counter[Id] = Counter(
{tid: len(all_scores[tid])
for tid in all_scores})
# Basic output statistics
read_seqs: int = sum(matchings.values())
if read_seqs == 0:
raise Exception(
red('\nERROR! ') + f'Cannot read any sequence from"{output_file}"')
filt_seqs: int = sum([len(scores) for scores in all_scores.values()])
if filt_seqs == 0:
raise Exception(red('\nERROR! ') + 'No sequence passed the filter!')
stat: SampleStats = SampleStats(minscore=minscore,
nt_read=nt_read,
scores=all_scores,
lens=all_length,
seq_read=read_seqs,
seq_filt=filt_seqs,
seq_clas=matchings[Match.DIRECT] +
matchings[Match.MULTI])
output.write(
gray(' Seqs read: ') + f'{stat.seq.read:_d}\t' + gray('[') +
f'{stat.nt_read}' + gray(']\n'))
output.write(
gray(' Seqs clas: ') + f'{stat.seq.clas:_d}\t' + gray('(') +
f'{stat.get_unclas_ratio():.2%}' + gray(' unclassified)\n'))
output.write(
gray(' Seqs pass: '******'{stat.seq.filt:_d}\t' + gray('(') +
f'{stat.get_reject_ratio():.2%}' + gray(' rejected)\n'))
multi_rel: float = matchings[Match.MULTI] / read_seqs
direct_rel: float = matchings[Match.DIRECT] / read_seqs
nodbhits_rel: float = matchings[Match.NODBHITS] / read_seqs
tooshort_rel: float = matchings[Match.READTOOSHORT] / read_seqs
lowscore_rel: float = matchings[Match.LOWSCORE] / read_seqs
output.write(f'\033[90m DB Matching: '
f'Multi =\033[0m {multi_rel:.1%}\033[90m '
f'Direct =\033[0m {direct_rel:.1%}\033[90m '
f'ReadTooShort =\033[0m {tooshort_rel:.1%}\033[90m '
f'LowScore =\033[0m {lowscore_rel:.1%}\033[90m '
f'NoDbHits =\033[0m {nodbhits_rel:.1%}\033[90m\n')
output.write(
gray(' Scores: min = ') + f'{stat.sco.mini:.1f},' + gray(' max = ') +
f'{stat.sco.maxi:.1f},' + gray(' avr = ') + f'{stat.sco.mean:.1f}\n')
output.write(
gray(' Length: min = ') + f'{stat.len.mini},' + gray(' max = ') +
f'{stat.len.maxi},' + gray(' avr = ') + f'{stat.len.mean}\n')
output.write(f' {stat.num_taxa}' + gray(f' taxa with assigned reads\n'))
# Select score output
out_scores: Dict[Id, Score]
if scoring is Scoring.LMAT:
out_scores = {tid: Score(mean(all_scores[tid])) for tid in all_scores}
else:
print(red('ERROR!'), f' LMAT: Unsupported Scoring "{scoring}"')
raise Exception('Unsupported scoring') # Return
return output.getvalue(), stat, abundances, out_scores
def main():
"""Main entry point to script."""
# Argument Parser Configuration
parser = argparse.ArgumentParser(
description='Extract reads following Centrifuge/Kraken output',
epilog=f'%(prog)s - {__author__} - {__date__}')
parser.add_argument('-V',
'--version',
action='version',
version=f'%(prog)s release {__version__} ({__date__})')
parser.add_argument('-f',
'--file',
action='store',
metavar='FILE',
required=True,
help='Centrifuge output file.')
parser.add_argument('-l',
'--limit',
action='store',
metavar='NUMBER',
type=int,
default=None,
help=('Limit of FASTQ reads to extract. '
'Default: no limit'))
parser.add_argument(
'-m',
'--maxreads',
action='store',
metavar='NUMBER',
type=int,
default=None,
help=('Maximum number of FASTQ reads to search for the taxa. '
'Default: no maximum'))
parser.add_argument(
'-n',
'--nodespath',
action='store',
metavar='PATH',
default=TAXDUMP_PATH,
help=('path for the nodes information files (nodes.dmp and names.dmp' +
' from NCBI'))
parser.add_argument(
'-i',
'--include',
action='append',
metavar='TAXID',
type=TaxId,
default=[],
help=('NCBI taxid code to include a taxon and all underneath ' +
'(multiple -i is available to include several taxid). ' +
'By default all the taxa is considered for inclusion.'))
parser.add_argument(
'-x',
'--exclude',
action='append',
metavar='TAXID',
type=TaxId,
default=[],
help=('NCBI taxid code to exclude a taxon and all underneath ' +
'(multiple -x is available to exclude several taxid)'))
parser.add_argument(
'-y',
'--minscore',
action='store',
metavar='NUMBER',
type=lambda txt: Score(float(txt)),
default=None,
help=('minimum score/confidence of the classification of a read '
'to pass the quality filter; all pass by default'))
filein = parser.add_mutually_exclusive_group(required=True)
filein.add_argument('-q',
'--fastq',
action='store',
metavar='FILE',
default=None,
help='Single FASTQ file (no paired-ends)')
filein.add_argument('-1',
'--mate1',
action='store',
metavar='FILE',
default=None,
help='Paired-ends FASTQ file for mate 1s '
'(filename usually includes _1)')
parser.add_argument('-2',
'--mate2',
action='store',
metavar='FILE',
default=None,
help='Paired-ends FASTQ file for mate 2s '
'(filename usually includes _2)')
# timing initialization
start_time: float = time.time()
# Program header
print(f'\n=-= {sys.argv[0]} =-= v{__version__} =-= {__date__} =-=\n')
sys.stdout.flush()
# Parse arguments
args = parser.parse_args()
output_file = args.file
nodesfile: Filename = Filename(os.path.join(args.nodespath, NODES_FILE))
namesfile: Filename = Filename(os.path.join(args.nodespath, NAMES_FILE))
excluding: Set[TaxId] = set(args.exclude)
including: Set[TaxId] = set(args.include)
fastq_1: Filename
fastq_2: Filename = args.mate2
if not fastq_2:
fastq_1 = args.fastq
else:
fastq_1 = args.mate1
# Load NCBI nodes, names and build children
plasmidfile: Filename = None
ncbi: Taxonomy = Taxonomy(nodesfile, namesfile, plasmidfile, False,
excluding, including)
# Build taxonomy tree
print(gray('Building taxonomy tree...'), end='')
sys.stdout.flush()
tree = TaxTree()
tree.grow(taxonomy=ncbi, look_ancestors=False)
print(green(' OK!'))
# Get the taxa
print(gray('Filtering taxa...'), end='')
sys.stdout.flush()
ranks: Ranks = Ranks({})
tree.get_taxa(ranks=ranks, include=including, exclude=excluding)
print(green(' OK!'))
taxids: Set[TaxId] = set(ranks)
taxlevels: TaxLevels = Rank.ranks_to_taxlevels(ranks)
num_taxlevels = Counter({rank: len(taxlevels[rank]) for rank in taxlevels})
num_taxlevels = +num_taxlevels
# Statistics about including taxa
print(f' {len(taxids)}\033[90m taxid selected in \033[0m', end='')
print(f'{len(num_taxlevels)}\033[90m different taxonomical levels:\033[0m')
for rank in num_taxlevels:
print(f' Number of different {rank}: {num_taxlevels[rank]}')
assert taxids, red('ERROR! No taxids to search for!')
# Get the records
records: List[SeqRecord] = []
num_seqs: int = 0
# timing initialization
start_time_load: float = time.perf_counter()
print(gray(f'Loading output file {output_file}...'), end='')
sys.stdout.flush()
try:
with open(output_file, 'rU') as file:
file.readline() # discard header
for num_seqs, record in enumerate(SeqIO.parse(file, 'centrifuge')):
tid: TaxId = record.annotations['taxID']
if tid not in taxids:
continue # Ignore read if low confidence
score: Score = Score(record.annotations['score'])
if args.minscore is not None and score < args.minscore:
continue
records.append(record)
except FileNotFoundError:
raise Exception(red('ERROR!') + 'Cannot read "' + output_file + '"')
print(green(' OK!'))
# Basic records statistics
print(
gray(' Load elapsed time: ') +
f'{time.perf_counter() - start_time_load:.3g}' + gray(' sec'))
print(f' \033[90mMatching reads: \033[0m{len(records):_d} \033[90m\t'
f'(\033[0m{len(records)/num_seqs:.4%}\033[90m of sample)')
sys.stdout.flush()
# FASTQ sequence dealing
# records_ids: List[SeqRecord] = [record.id for record in records]
records_ids: Set[SeqRecord] = {record.id for record in records}
seqs1: List[SeqRecord] = []
seqs2: List[SeqRecord] = []
extracted: int = 0
i: int = 0
if fastq_2:
print(
f'\033[90mLoading FASTQ files {fastq_1} and {fastq_2}...\n'
f'Mseqs: \033[0m',
end='')
sys.stdout.flush()
try:
with open(fastq_1, 'rU') as file1, open(fastq_2, 'rU') as file2:
for i, (rec1, rec2) in enumerate(
zip(SeqIO.parse(file1, 'quickfastq'),
SeqIO.parse(file2, 'quickfastq'))):
if not records_ids or (args.maxreads and i >= args.maxreads
) or (args.limit
and extracted >= args.limit):
break
elif not i % 1000000:
print(f'{i//1000000:_d}', end='')
sys.stdout.flush()
elif not i % 100000:
print('.', end='')
sys.stdout.flush()
try:
records_ids.remove(rec1.id)
except KeyError:
pass
else:
seqs1.append(rec1)
seqs2.append(rec2)
extracted += 1
except FileNotFoundError:
raise Exception('\n\033[91mERROR!\033[0m Cannot read FASTQ files')
else:
print(f'\033[90mLoading FASTQ files {fastq_1}...\n'
f'Mseqs: \033[0m',
end='')
sys.stdout.flush()
try:
with open(fastq_1, 'rU') as file1:
for i, rec1 in enumerate(SeqIO.parse(file1, 'quickfastq')):
if not records_ids or (args.maxreads and i >= args.maxreads
) or (args.limit
and extracted >= args.limit):
break
elif not i % 1000000:
print(f'{i//1000000:_d}', end='')
sys.stdout.flush()
elif not i % 100000:
print('.', end='')
sys.stdout.flush()
try:
records_ids.remove(rec1.id)
except KeyError:
pass
else:
seqs1.append(rec1)
extracted += 1
except FileNotFoundError:
raise Exception('\n\033[91mERROR!\033[0m Cannot read FASTQ file')
print(cyan(f' {i/1e+6:.3g} Mseqs'), green('OK! '))
def format_filename(fastq: Filename) -> Filename:
"""Auxiliary function to properly format the output filenames.
Args:
fastq: Complete filename of the fastq input file
Returns: Filename of the rextracted fastq output file
"""
fastq_filename, _ = os.path.splitext(fastq)
output_list: List[str] = [fastq_filename, '_rxtr']
if including:
output_list.append('_incl')
output_list.extend('_'.join(including))
if excluding:
output_list.append('_excl')
output_list.extend('_'.join(excluding))
output_list.append('.fastq')
return Filename(''.join(output_list))
filename1: Filename = format_filename(fastq_1)
SeqIO.write(seqs1, filename1, 'quickfastq')
print(gray('Wrote'), magenta(f'{len(seqs1)}'), gray('reads in'), filename1)
if fastq_2:
filename2: Filename = format_filename(fastq_2)
SeqIO.write(seqs2, filename2, 'quickfastq')
print(gray('Wrote'), magenta(f'{len(seqs1)}'), gray('reads in'),
filename2)
# Timing results
print(gray('Total elapsed time:'),
time.strftime("%H:%M:%S", time.gmtime(time.time() - start_time)))
def read_output(
output_file: Filename,
scoring: Scoring = Scoring.SHEL,
minscore: Score = None,
) -> Tuple[str, SampleStats, Counter[TaxId], Dict[TaxId, Score]]:
"""
Read Centrifuge output file
Args:
output_file: output file name
scoring: type of scoring to be applied (see Scoring class)
minscore: minimum confidence level for the classification
Returns:
log string, statistics, abundances counter, scores dict
"""
output: io.StringIO = io.StringIO(newline='')
all_scores: Dict[TaxId, List[Score]] = {}
all_length: Dict[TaxId, List[int]] = {}
num_read: int = 0
nt_read: int = 0
num_uncl: int = 0
error_read: int = None
output.write(gray(f'Loading output file {output_file}... '))
try:
with open(output_file, 'r') as file:
file.readline() # discard header
for output_line in file:
try:
_, _, _tid, _score, _, _, _length, *_ = output_line.split(
'\t')
except ValueError:
print(
red('Error'), f'parsing line: ({output_line}) '
f'in {output_file}. Ignoring line!')
error_read = num_read + 1
continue
tid = TaxId(_tid)
try:
# From Centrifuge score get "single hit equivalent length"
shel = Score(float(_score)**0.5 + 15)
length = int(_length)
except ValueError:
print(red('Error'), f'parsing score ({_score}) for query',
f'length ({_length}) for taxid {_tid}',
f'in {output_file}. Ignoring line!')
continue
num_read += 1
nt_read += length
if tid == UNCLASSIFIED: # Just count unclassified reads
num_uncl += 1
continue
elif minscore is not None and shel < minscore:
continue # Ignore read if low confidence
try:
all_scores[tid].append(shel)
except KeyError:
all_scores[tid] = [
shel,
]
try:
all_length[tid].append(length)
except KeyError:
all_length[tid] = [
length,
]
except FileNotFoundError:
raise Exception(red('\nERROR! ') + f'Cannot read "{output_file}"')
if error_read == num_read + 1: # Check if error in last line: truncated!
print(yellow('Warning!'), f'{output_file} seems truncated!')
counts: Counter[TaxId] = Counter(
{tid: len(all_scores[tid])
for tid in all_scores})
output.write(green('OK!\n'))
if num_read == 0:
raise Exception(
red('\nERROR! ') + f'Cannot read any sequence from"{output_file}"')
filt_seqs: int = sum([len(scores) for scores in all_scores.values()])
if filt_seqs == 0:
raise Exception(red('\nERROR! ') + 'No sequence passed the filter!')
# Get statistics
stat: SampleStats = SampleStats(minscore=minscore,
nt_read=nt_read,
scores=all_scores,
lens=all_length,
seq_read=num_read,
seq_unclas=num_uncl,
seq_filt=filt_seqs)
# Output statistics
output.write(
gray(' Seqs read: ') + f'{stat.seq.read:_d}\t' + gray('[') +
f'{stat.nt_read}' + gray(']\n'))
output.write(
gray(' Seqs clas: ') + f'{stat.seq.clas:_d}\t' + gray('(') +
f'{stat.get_unclas_ratio():.2%}' + gray(' unclassified)\n'))
output.write(
gray(' Seqs pass: '******'{stat.seq.filt:_d}\t' + gray('(') +
f'{stat.get_reject_ratio():.2%}' + gray(' rejected)\n'))
output.write(
gray(' Scores: min = ') + f'{stat.sco.mini:.1f},' + gray(' max = ') +
f'{stat.sco.maxi:.1f},' + gray(' avr = ') + f'{stat.sco.mean:.1f}\n')
output.write(
gray(' Length: min = ') + f'{stat.len.mini},' + gray(' max = ') +
f'{stat.len.maxi},' + gray(' avr = ') + f'{stat.len.mean}\n')
output.write(f' {stat.num_taxa}' + gray(f' taxa with assigned reads\n'))
# Select score output
out_scores: Dict[TaxId, Score]
if scoring is Scoring.SHEL:
out_scores = {tid: Score(mean(all_scores[tid])) for tid in all_scores}
elif scoring is Scoring.LENGTH:
out_scores = {tid: Score(mean(all_length[tid])) for tid in all_length}
elif scoring is Scoring.LOGLENGTH:
out_scores = {
tid: Score(log10(mean(all_length[tid])))
for tid in all_length
}
elif scoring is Scoring.NORMA:
scores: Dict[TaxId, Score] = {
tid: Score(mean(all_scores[tid]))
for tid in all_scores
}
lengths: Dict[TaxId, Score] = {
tid: Score(mean(all_length[tid]))
for tid in all_length
}
out_scores = {
tid: Score(scores[tid] / lengths[tid] * 100)
for tid in scores
}
else:
raise Exception(f'\n\033[91mERROR!\033[0m Unknown Scoring "{scoring}"')
# Return
return output.getvalue(), stat, counts, out_scores
def configure_parser():
"""Argument Parser Configuration"""
parser = argparse.ArgumentParser(
description='Analyze results of metagenomic taxonomic classifiers',
epilog=f'%(prog)s - Release {__version__} - {__date__}' + LICENSE,
formatter_class=argparse.RawDescriptionHelpFormatter)
parser.add_argument(
'-V',
'--version',
action='version',
version=f'%(prog)s version {__version__} released in {__date__}')
parser_in = parser.add_argument_group(
'input', 'Define Recentrifuge input files and formats')
parser_in.add_argument('-n',
'--nodespath',
action='store',
metavar='PATH',
default=TAXDUMP_PATH,
help=('path for the nodes information files '
'(nodes.dmp and names.dmp from NCBI)'))
parser_filein = parser_in.add_mutually_exclusive_group(required=True)
parser_filein.add_argument(
'-f',
'--file',
action='append',
metavar='FILE',
type=Filename,
help=('Centrifuge output files. If a single directory is entered, '
'every .out file inside will be taken as a different sample.'
' Multiple -f is available to include several samples.'))
parser_filein.add_argument(
'-l',
'--lmat',
action='append',
metavar='FILE',
type=Filename,
default=None,
help=('LMAT output dir or file prefix. If just "." is entered, '
'every subdirectory under the current directory will be '
'taken as a sample and scanned looking for LMAT output files'
'. Multiple -l is available to include several samples.'))
parser_filein.add_argument(
'-k',
'--clark',
action='append',
metavar='FILE',
type=Filename,
help=('CLARK(S) output files. If a single directory is entered, '
'every .csv file inside will be taken as a different sample.'
' Multiple -k is available to include several samples.'))
parser_filein.add_argument(
'-r',
'--report',
action='append',
metavar='FILE',
type=Filename,
help=('Centrifuge/Kraken report files '
'(multiple -r is available to include several samples)'))
parser_out = parser.add_argument_group(
'output', 'Related to the Recentrifuge output files')
parser_out.add_argument(
'-o',
'--outhtml',
action='store',
metavar='FILE',
type=Filename,
help='HTML output file (if not given, the filename will be '
'inferred from input files)')
parser_out.add_argument(
'-e',
'--excel',
action='store',
metavar='OUTPUT_TYPE',
choices=[str(excel) for excel in Excel],
default=str(Excel(0)),
help=(f'type of excel report to be generated, and can be one of '
f'{[str(excel) for excel in Excel]}'))
parser_coarse = parser.add_argument_group(
'tuning', 'Coarse tuning of algorithm parameters')
parser_cross = parser_coarse.add_mutually_exclusive_group(
required=False)
parser_cross.add_argument(
'-c',
'--controls',
action='store',
metavar='CONTROLS_NUMBER',
type=int,
default=0,
help=('this number of first samples will be treated as negative '
'controls; default is no controls'))
parser_coarse.add_argument(
'-s',
'--scoring',
action='store',
metavar='SCORING',
choices=[str(each_score) for each_score in Scoring],
default=str(Scoring(0)),
help=(f'type of scoring to be applied, and can be one of '
f'{[str(scoring) for scoring in Scoring]}'))
parser_coarse.add_argument(
'-y',
'--minscore',
action='store',
metavar='NUMBER',
type=lambda txt: Score(float(txt)),
default=None,
help=('minimum score/confidence of the classification of a read '
'to pass the quality filter; all pass by default'))
parser_coarse.add_argument(
'-m',
'--mintaxa',
action='store',
metavar='INT',
type=int,
default=DEFMINTAXA,
help='minimum taxa to avoid collapsing one level to the parent one'
)
parser_coarse.add_argument(
'-x',
'--exclude',
action='append',
metavar='TAXID',
type=Id,
default=[],
help=('NCBI taxid code to exclude a taxon and all underneath '
'(multiple -x is available to exclude several taxid)'))
parser_coarse.add_argument(
'-i',
'--include',
action='append',
metavar='TAXID',
type=Id,
default=[],
help=('NCBI taxid code to include a taxon and all underneath '
'(multiple -i is available to include several taxid); '
'by default, all the taxa are considered for inclusion'))
parser_cross.add_argument('-a',
'--avoidcross',
action='store_true',
help='avoid cross analysis')
parser_fine = parser.add_argument_group(
'fine tuning', 'Fine tuning of algorithm parameters')
parser_fine.add_argument(
'-z',
'--ctrlminscore',
action='store',
metavar='NUMBER',
type=lambda txt: Score(float(txt)),
default=None,
help=('minimum score/confidence of the classification of a read '
'in control samples to pass the quality filter; if defaults '
'to "minscore"'))
parser_fine.add_argument(
'-w',
'--ctrlmintaxa',
action='store',
metavar='INT',
type=int,
default=None,
help='minimum taxa to avoid collapsing one level to the parent one'
' in control samples; it defaults to "mintaxa"')
parser_fine.add_argument(
'-u',
'--summary',
action='store',
metavar='OPTION',
choices=['add', 'only', 'avoid'],
default='add',
help=(
'select to "add" summary samples to other samples, or to '
'"only" show summary samples or to "avoid" summaries at all'))
parser_fine.add_argument(
'-t',
'--takeoutroot',
action='store_true',
help='remove counts directly assigned to the "root" level')
parser_fine.add_argument('--nokollapse',
action='store_true',
help='show the "cellular organisms" taxon')
parser_mode = parser.add_argument_group('advanced',
'Advanced modes of running')
parser_mode.add_argument(
'--dummy', # hidden flag: just generate a dummy plot for JS debug
action='store_true',
help=argparse.SUPPRESS)
parser_mode.add_argument(
'-g',
'--debug',
action='store_true',
help='increase output verbosity and perform additional checks')
parser_mode.add_argument('--sequential',
action='store_true',
help='deactivate parallel processing')
return parser
def process_output(
*args,
**kwargs) -> Tuple[Sample, TaxTree, SampleDataById, SampleStats, Err]:
"""
Process classifiers output files (to be usually called in parallel!).
"""
# timing initialization
start_time: float = time.perf_counter()
# Recover input and parameters
target_file: Filename = args[0]
debug: bool = kwargs['debug']
is_ctrl: bool = args[1]
if debug:
print(gray('Processing'), blue('ctrl' if is_ctrl else 'sample'),
target_file, gray('...'))
sys.stdout.flush()
ontology: Ontology = kwargs['ontology']
mintaxa: Optional[int] = (kwargs['ctrlmintaxa']
if is_ctrl else kwargs['mintaxa'])
minscore: Score = kwargs['ctrlminscore'] if is_ctrl else kwargs['minscore']
including: Union[Tuple, Set[Id]] = ontology.including
excluding: Union[Tuple, Set[Id]] = ontology.excluding
scoring: Scoring = kwargs['scoring']
classifier: Classifier = kwargs['classifier']
genfmt: GenericFormat = kwargs['genfmt']
output: io.StringIO = io.StringIO(newline='')
def vwrite(*args):
"""Print only if verbose/debug mode is enabled"""
if kwargs['debug']:
output.write(' '.join(str(item) for item in args))
sample: Sample = Sample(os.path.splitext(target_file)[0])
error: Err = Err.NO_ERROR
# Read taxonomic classifier output files to get abundances
read_method: Callable[ # Format: [[Input], Output]
[Filename, Scoring, Optional[Score]],
Tuple[str, SampleStats, Counter[Id], Dict[Id, Score]]]
log: str
stat: SampleStats
counts: Counter[Id]
scores: Dict[Id, Score]
if classifier is Classifier.GENERIC: # Direct call to generic method
log, stat, counts, scores = read_generic_output(
target_file, scoring, minscore, genfmt)
else: # Use read_method
if classifier is Classifier.KRAKEN:
read_method = read_kraken_output
elif classifier is Classifier.CLARK:
read_method = read_clark_output
elif classifier is Classifier.LMAT:
read_method = read_lmat_output
elif classifier is Classifier.CENTRIFUGE:
read_method = read_output
else:
raise Exception(red('\nERROR!'),
f'taxclass: Unknown classifier "{classifier}".')
log, stat, counts, scores = read_method(target_file, scoring, minscore)
output.write(log)
# Complete/Update fields in stats
stat.is_ctrl = is_ctrl # set control nature of the sample
if mintaxa is not None: # manual mintaxa has precedence over automatic
stat.mintaxa = mintaxa
else: # update local value with the automatically guessed value
mintaxa = stat.mintaxa
# Move cellular_organisms counts to root, in case
if ontology.collapse and counts[CELLULAR_ORGANISMS]:
vwrite(gray('Moving'), counts[CELLULAR_ORGANISMS],
gray('"CELLULAR_ORGANISMS" reads to "ROOT"... \n'))
if counts[ontology.ROOT]:
stat.decrease_filtered_taxids()
scores[ontology.ROOT] = Score(
(scores[CELLULAR_ORGANISMS] * counts[CELLULAR_ORGANISMS] +
scores[ontology.ROOT] * counts[ontology.ROOT]) /
(counts[CELLULAR_ORGANISMS] + counts[ontology.ROOT]))
else:
scores[ontology.ROOT] = scores[CELLULAR_ORGANISMS]
counts[ontology.ROOT] += counts[CELLULAR_ORGANISMS]
counts[CELLULAR_ORGANISMS] = 0
scores[CELLULAR_ORGANISMS] = NO_SCORE
# Remove root counts, in case
if kwargs['root'] and counts[ontology.ROOT]:
vwrite(gray('Removing'), counts[ontology.ROOT],
gray('"ROOT" reads... '))
stat.seq = stat.seq._replace(filt=stat.seq.filt -
counts[ontology.ROOT])
stat.decrease_filtered_taxids()
counts[ontology.ROOT] = 0
scores[ontology.ROOT] = NO_SCORE
vwrite(green('OK!'), '\n')
# Building ontology tree
output.write(
gray('Building from raw data with mintaxa = ') + f'{mintaxa:_d}' +
gray(' ... \n'))
vwrite(gray(' Building ontology tree with all-in-1... '))
tree = TaxTree()
ancestors: Set[Id]
orphans: Set[Id]
ancestors, orphans = ontology.get_ancestors(counts.keys())
out = SampleDataById(['all'])
tree.allin1(ontology=ontology,
counts=counts,
scores=scores,
ancestors=ancestors,
min_taxa=mintaxa,
include=including,
exclude=excluding,
out=out)
out.purge_counters()
vwrite(green('OK!'), '\n')
# Stats: Complete final value for TaxIDs after tree building and folding
final_taxids: int = len(out.counts) if out.counts is not None else 0
stat.set_final_taxids(final_taxids)
# Check for additional loss of reads (due to include/exclude an orphans)
output.write(gray(' Check for more seqs lost ([in/ex]clude affects)... '))
if out.counts is not None:
discard: int = sum(counts.values()) - sum(out.counts.values())
if discard:
output.write(
blue('\n Info:') + f' {discard} ' +
gray('additional seqs discarded (') +
f'{discard/sum(counts.values()):.3%} ' +
gray('of accepted)\n'))
else:
output.write(green('OK!\n'))
else:
output.write(red('No counts in sample tree!\n'))
# Warn or give detailed stats about orphan taxid and orphan seqs
if debug:
vwrite(gray(' Checking taxid loss (orphans)... '))
lost: int = 0
if orphans:
for orphan in orphans:
vwrite(yellow(' Warning!'), gray('Orphan taxid'),
f'{orphan}\n')
lost += counts[orphan]
vwrite(
yellow(' WARNING!'), f'{len(orphans)} orphan taxids ('
f'{len(orphans)/len(counts):.2%} of accepted)\n'
f' and {lost} orphan sequences ('
f'{lost/sum(counts.values()):.3%} of accepted)\n')
else:
vwrite(green('OK!\n'))
elif orphans:
output.write(
yellow('\n Warning!') + f' {len(orphans)} orphan taxids' +
gray(' (rerun with --debug for details)\n'))
# Check the removal of TaxIDs (accumulation of leaves in parents)
if debug and not excluding and including == {ontology.ROOT}:
vwrite(gray(' Assess accumulation due to "folding the tree"...\n'))
migrated: int = 0
if out.counts is not None:
for taxid in counts:
if out.counts[taxid] == 0:
migrated += 1
vwrite(
blue(' Info:'),
gray(f'Folded TaxID {taxid} (') +
f'{ontology.get_name(taxid)}' + gray(') with ') +
f'{counts[taxid]}' + gray(' original seqs\n'))
if migrated:
vwrite(
blue(' INFO:'), f'{migrated} TaxIDs folded ('
f'{migrated/len(+counts):.2%} of TAF —TaxIDs after filtering—)'
'\n')
vwrite(
blue(' INFO:'), f'Final assigned TaxIDs: {final_taxids} '
f'(reduced to {final_taxids/len(+counts):.2%} of '
'number of TAF)\n')
else:
vwrite(blue(' INFO:'), gray('No migration!'), green('OK!\n'))
# Print last message and check if the sample is void
if out.counts:
output.write(sample + blue(' ctrl ' if is_ctrl else ' sample ') +
green('OK!\n'))
elif is_ctrl:
output.write(sample + red(' ctrl VOID!\n'))
error = Err.VOID_CTRL
else:
output.write(sample + blue(' sample ') + yellow('VOID\n'))
error = Err.VOID_SAMPLE
# Timing results
output.write(
gray('Load elapsed time: ') +
f'{time.perf_counter() - start_time:.3g}' + gray(' sec\n'))
print(output.getvalue())
sys.stdout.flush()
return sample, tree, out, stat, error
def read_kraken_output(
output_file: Filename,
scoring: Scoring = Scoring.KRAKEN,
minscore: Score = None,
) -> Tuple[str, SampleStats, Counter[Id], Dict[Id, Score]]:
"""
Read Kraken output file
Args:
output_file: output file name
scoring: type of scoring to be applied (see Scoring class)
minscore: minimum confidence level for the classification
Returns:
log string, statistics, abundances counter, scores dict
"""
output: io.StringIO = io.StringIO(newline='')
all_scores: Dict[Id, List[Score]] = {}
all_kmerel: Dict[Id, List[Score]] = {}
all_length: Dict[Id, List[int]] = {}
taxids: Set[Id] = set()
num_read: int = 0
nt_read: int = 0
num_uncl: int = 0
last_error_read: int = -1 # Number of read of the last error
num_errors: int = 0 # Number or reads discarded due to error
output.write(gray(f'Loading output file {output_file}... '))
try:
with open(output_file, 'r') as file:
# Check number of cols in header
header = file.readline().split('\t')
if len(header) != 5:
print(
red('\nERROR! ') + 'Kraken output format of ',
yellow(f'"{output_file}"'), 'not supported.')
print(magenta('Expected:'),
'C/U, ID, taxid, length, list of mappings')
print(magenta('Found:'), '\t'.join(header), end='')
print(blue('HINT:'), 'Use Kraken or Kraken2 direct output.')
raise Exception('Unsupported file format. Aborting.')
for raw_line in file:
try:
output_line = raw_line.strip()
(_clas, _label, _tid, _length,
_maps) = output_line.split('\t')
except ValueError:
print(
yellow('Failure'), 'parsing line elements:'
f' {output_line} in {output_file}'
'. Ignoring line!')
last_error_read = num_read + 1
num_errors += 1
continue
try:
length: int = sum(map(int, _length.split('|')))
num_read += 1
nt_read += length
if _clas == UNCLASSIFIED: # Just count unclassified reads
num_uncl += 1
continue
tid: Id = Id(_tid)
maps: List[str] = _maps.split()
try:
maps.remove('|:|')
except ValueError:
pass
mappings: Counter[Id] = col.Counter()
for pair in maps:
couple: List[str] = pair.split(':')
mappings[Id(couple[0])] += int(couple[1])
# From Kraken score get "single hit equivalent length"
shel: Score = Score(mappings[tid] + K_MER_SIZE)
score: Score = Score(mappings[tid] /
sum(mappings.values()) *
100) # % relative to all k-mers
except ValueError:
print(
yellow('Failure'), 'parsing line elements:'
f' {output_line} in {output_file}'
'. Ignoring line!')
last_error_read = num_read + 1
num_errors += 1
continue
else:
taxids.add(tid) # Save all the tids of classified reads
if minscore is not None: # Decide if ignore read if low score
if scoring is Scoring.KRAKEN:
if score < minscore:
continue
else:
if shel < minscore:
continue
try:
all_scores[tid].append(shel)
except KeyError:
all_scores[tid] = [
shel,
]
try:
all_kmerel[tid].append(score)
except KeyError:
all_kmerel[tid] = [
score,
]
try:
all_length[tid].append(length)
except KeyError:
all_length[tid] = [
length,
]
except FileNotFoundError:
raise Exception(red('\nERROR! ') + f'Cannot read "{output_file}"')
if last_error_read == num_read + 1: # Check error in last line: truncated!
print(yellow('Warning!'), f'{output_file} seems truncated!')
counts: Counter[Id] = col.Counter(
{tid: len(all_scores[tid])
for tid in all_scores})
output.write(green('OK!\n'))
if num_read == 0:
raise Exception(
red('\nERROR! ') +
f'Cannot read any sequence from "{output_file}"')
filt_seqs: int = sum([len(scores) for scores in all_scores.values()])
if filt_seqs == 0:
raise Exception(red('\nERROR! ') + 'No sequence passed the filter!')
# Get statistics
stat: SampleStats = SampleStats(minscore=minscore,
nt_read=nt_read,
lens=all_length,
scores=all_scores,
scores2=all_kmerel,
seq_read=num_read,
seq_unclas=num_uncl,
seq_filt=filt_seqs,
tid_clas=len(taxids))
# Output statistics
if num_errors:
output.write(
gray(' Seqs fail: ') + red(f'{num_errors:_d}\t') +
gray('(Last error in read ') + red(f'{last_error_read}') +
gray(')\n'))
output.write(
gray(' Seqs read: ') + f'{stat.seq.read:_d}\t' + gray('[') +
f'{stat.nt_read}' + gray(']\n'))
output.write(
gray(' Seqs clas: ') + f'{stat.seq.clas:_d}\t' + gray('(') +
f'{stat.get_unclas_ratio():.2%}' + gray(' unclassified)\n'))
output.write(
gray(' Seqs pass: '******'{stat.seq.filt:_d}\t' + gray('(') +
f'{stat.get_reject_ratio():.2%}' + gray(' rejected)\n'))
output.write(
gray(' Scores SHEL: min = ') + f'{stat.sco.mini:.1f},' +
gray(' max = ') + f'{stat.sco.maxi:.1f},' + gray(' avr = ') +
f'{stat.sco.mean:.1f}\n')
output.write(
gray(' Coverage(%): min = ') + f'{stat.sco2.mini:.1f},' +
gray(' max = ') + f'{stat.sco2.maxi:.1f},' + gray(' avr = ') +
f'{stat.sco2.mean:.1f}\n')
output.write(
gray(' Read length: min = ') + f'{stat.len.mini},' + gray(' max = ') +
f'{stat.len.maxi},' + gray(' avr = ') + f'{stat.len.mean}\n')
output.write(
gray(' TaxIds: by classifier = ') + f'{stat.tid.clas}' +
gray(', by filter = ') + f'{stat.tid.filt}\n')
# Select score output
out_scores: Dict[Id, Score]
if scoring is Scoring.SHEL:
out_scores = {tid: Score(mean(all_scores[tid])) for tid in all_scores}
elif scoring is Scoring.KRAKEN:
out_scores = {tid: Score(mean(all_kmerel[tid])) for tid in all_kmerel}
elif scoring is Scoring.LENGTH:
out_scores = {tid: Score(mean(all_length[tid])) for tid in all_length}
elif scoring is Scoring.LOGLENGTH:
out_scores = {
tid: Score(log10(mean(all_length[tid])))
for tid in all_length
}
elif scoring is Scoring.NORMA:
scores: Dict[Id, Score] = {
tid: Score(mean(all_scores[tid]))
for tid in all_scores
}
lengths: Dict[Id, Score] = {
tid: Score(mean(all_length[tid]))
for tid in all_length
}
out_scores = {
tid: Score(scores[tid] / lengths[tid] * 100)
for tid in scores
}
else:
print(red('ERROR!'), f'kraken: Unsupported Scoring "{scoring}"')
raise Exception('Unsupported scoring')
# Return
return output.getvalue(), stat, counts, out_scores
def read_generic_output(
output_file: Filename,
scoring: Scoring = Scoring.GENERIC,
minscore: Score = None,
genfmt: GenericFormat = None
) -> Tuple[str, SampleStats, Counter[Id], Dict[Id, Score]]:
"""
Read an output file from a generic classifier
Args:
output_file: output file name
scoring: type of scoring to be applied (see Scoring class)
minscore: minimum confidence level for the classification
genfmt: GenericFormat object specifying the files format
Returns:
log string, statistics, abundances counter, scores dict
"""
# Initialization of variables
output: io.StringIO = io.StringIO(newline='')
all_scores: Dict[Id, List[Score]] = {}
all_length: Dict[Id, List[int]] = {}
taxids: Set[Id] = set()
num_read: int = 0
nt_read: int = 0
num_uncl: int = 0
last_error_read: int = -1 # Number of read of the last error
num_errors: int = 0 # Number or reads discarded due to error
output.write(gray(f'Loading output file {output_file}... '))
# Check format
if not isinstance(genfmt, GenericFormat):
raise Exception(
red('\nERROR!'),
'Missing GenericFormat when reading a generic output.')
try:
with open(output_file, 'r') as file:
# Main loop processing each file line
for raw_line in file:
raw_line = raw_line.strip(' \n\t')
splitting: str
if genfmt.typ is GenericType.CSV:
splitting = ','
elif genfmt.typ is GenericType.TSV:
splitting = '\t'
elif genfmt.typ is GenericType.SSV:
splitting = ' '
else:
raise Exception(f'ERROR! Unknown GenericType {genfmt.typ}')
output_line: List[str] = raw_line.split(splitting)
if len(output_line) < GenericFormat.MIN_COLS:
if num_read == 0 and last_error_read < 0:
last_error_read = 0
print(yellow('Warning!'), 'Skipping header of '
f'{output_file}')
continue # Not account for the header as an error
raise Exception(
red('\nERROR!') + ' Line ' + yellow(f'{output_line}') +
'\n\tin ' + yellow(f'{output_file}') + ' has < ' +
blue(f'{GenericFormat.MIN_COLS}') + ' required ' +
'columns.\n\tPlease check the file.')
try:
tid: Id = Id(output_line[genfmt.tid - 1].strip(' "'))
length: int = int(output_line[genfmt.len - 1].strip(' "'))
if tid == genfmt.unc: # Avoid read score for unclass reads
num_read += 1
nt_read += length
num_uncl += 1
continue
score: Score = Score(
float(output_line[genfmt.sco - 1].strip(' "')))
except ValueError:
if num_read == 0 and last_error_read < 0:
last_error_read = 0
print(yellow('Warning!'), 'Skipping header of '
f'{output_file}')
continue # Not account for the header as a failure
print(
yellow('Failure'), 'parsing line elements:'
f' {output_line} in {output_file}'
'. Ignoring line!')
last_error_read = num_read + 1
num_errors += 1
if num_read > 100 and num_errors > 0.5 * num_read:
print(
red('ERROR!'),
'Unreliable file processing: rate of problematic'
f' reads is {num_errors/num_read*100:_d}, beyond'
' 50%, after 100 reads. Please check the format '
f'of the file "{output_file}".')
raise
else:
continue
num_read += 1
nt_read += length
taxids.add(tid) # Save all the tids of classified reads
if minscore is not None and score < minscore:
continue # Discard read if low confidence
try:
all_scores[tid].append(score)
except KeyError:
all_scores[tid] = [
score,
]
try:
all_length[tid].append(length)
except KeyError:
all_length[tid] = [
length,
]
except FileNotFoundError:
raise Exception(red('\nERROR! ') + f'Cannot read "{output_file}"')
if last_error_read == num_read + 1: # Check error in last line: truncated!
print(yellow('Warning!'), f'{output_file} seems truncated!')
counts: Counter[Id] = col.Counter(
{tid: len(all_scores[tid])
for tid in all_scores})
output.write(green('OK!\n'))
if num_read == 0:
raise Exception(
red('\nERROR! ') +
f'Cannot read any sequence from "{output_file}"')
filt_seqs: int = sum([len(scores) for scores in all_scores.values()])
if filt_seqs == 0:
raise Exception(red('\nERROR! ') + 'No sequence passed the filter!')
# Get statistics
stat: SampleStats = SampleStats(minscore=minscore,
nt_read=nt_read,
lens=all_length,
scores=all_scores,
seq_read=num_read,
seq_unclas=num_uncl,
seq_filt=filt_seqs,
tid_clas=len(taxids))
# Output statistics
if num_errors:
output.write(
gray(' Seqs fail: ') + red(f'{num_errors:_d}\t') +
gray('(Last error in read ') + red(f'{last_error_read}') +
gray(')\n'))
output.write(
gray(' Seqs read: ') + f'{stat.seq.read:_d}\t' + gray('[') +
f'{stat.nt_read}' + gray(']\n'))
output.write(
gray(' Seqs clas: ') + f'{stat.seq.clas:_d}\t' + gray('(') +
f'{stat.get_unclas_ratio():.2%}' + gray(' unclassified)\n'))
output.write(
gray(' Seqs pass: '******'{stat.seq.filt:_d}\t' + gray('(') +
f'{stat.get_reject_ratio():.2%}' + gray(' rejected)\n'))
output.write(
gray(' Scores: min = ') + f'{stat.sco.mini:.1f},' + gray(' max = ') +
f'{stat.sco.maxi:.1f},' + gray(' avr = ') + f'{stat.sco.mean:.1f}\n')
output.write(
gray(' Read length: min = ') + f'{stat.len.mini},' + gray(' max = ') +
f'{stat.len.maxi},' + gray(' avr = ') + f'{stat.len.mean}\n')
output.write(
gray(' TaxIds: by classifier = ') + f'{stat.tid.clas}' +
gray(', by filter = ') + f'{stat.tid.filt}\n')
# Select score output
out_scores: Dict[Id, Score]
if scoring is Scoring.GENERIC:
out_scores = {tid: Score(mean(all_scores[tid])) for tid in all_scores}
elif scoring is Scoring.LENGTH:
out_scores = {tid: Score(mean(all_length[tid])) for tid in all_length}
elif scoring is Scoring.LOGLENGTH:
out_scores = {
tid: Score(log10(mean(all_length[tid])))
for tid in all_length
}
elif scoring is Scoring.NORMA:
scores: Dict[Id, Score] = {
tid: Score(mean(all_scores[tid]))
for tid in all_scores
}
lengths: Dict[Id, Score] = {
tid: Score(mean(all_length[tid]))
for tid in all_length
}
out_scores = {
tid: Score(scores[tid] / lengths[tid] * 100)
for tid in scores
}
else:
raise Exception(red('\nERROR!'),
f'Generic: Unsupported Scoring "{scoring}"')
# Return
return output.getvalue(), stat, counts, out_scores