diff options
Diffstat (limited to 'bh20sequploader/qc_fasta.py')
| -rw-r--r-- | bh20sequploader/qc_fasta.py | 73 |
1 files changed, 37 insertions, 36 deletions
diff --git a/bh20sequploader/qc_fasta.py b/bh20sequploader/qc_fasta.py index e198430..37eb4e8 100644 --- a/bh20sequploader/qc_fasta.py +++ b/bh20sequploader/qc_fasta.py @@ -25,7 +25,7 @@ def read_fasta(sequence): raise ValueError("FASTA file contains multiple entries") return label, bases -def qc_fasta(arg_sequence): +def qc_fasta(arg_sequence, check_with_clustalw=True): log.debug("Starting qc_fasta") schema_resource = pkg_resources.resource_stream(__name__, "validation/formats") with tempfile.NamedTemporaryFile() as tmp: @@ -51,47 +51,48 @@ def qc_fasta(arg_sequence): if seq_type == "text/fasta": # ensure that contains only one entry submitlabel, submitseq = read_fasta(sequence) + sequence.seek(0) + sequence.detach() - with tempfile.NamedTemporaryFile() as tmp1: - refstring = pkg_resources.resource_string(__name__, "SARS-CoV-2-reference.fasta") - tmp1.write(refstring) - tmp1.write(submitlabel.encode("utf8")) - tmp1.write(("".join(submitseq)).encode("utf8")) - tmp1.flush() - subbp = 0 - refbp = 0 - similarity = 0 - try: - cmd = ["clustalw", "-infile="+tmp1.name, - "-quicktree", "-iteration=none", "-type=DNA"] - print("QC checking similarity to reference") - print(" ".join(cmd)) - result = subprocess.run(cmd, stdout=subprocess.PIPE) - res = result.stdout.decode("utf-8") - g1 = re.search(r"^Sequence 1: [^ ]+ +(\d+) bp$", res, flags=re.MULTILINE) - refbp = float(g1.group(1)) - g2 = re.search(r"^Sequence 2: [^ ]+ +(\d+) bp$", res, flags=re.MULTILINE) - subbp = float(g2.group(1)) - g3 = re.search(r"^Sequences \(1:2\) Aligned\. Score: (\d+(\.\d+)?)$", res, flags=re.MULTILINE) - similarity = float(g3.group(1)) + if not check_with_clustalw: + return ("sequence.fasta"+gz, seqlabel) - print(g1.group(0)) - print(g2.group(0)) - print(g3.group(0)) - except Exception as e: - logging.warn("Error trying to QC against reference sequence using 'clustalw': %s", e) + with tempfile.NamedTemporaryFile() as tmp1: + with tempfile.NamedTemporaryFile() as tmp2: + refstring = pkg_resources.resource_string(__name__, "SARS-CoV-2-reference.fasta") + tmp1.write(refstring) + tmp1.flush() + tmp2.write(submitlabel.encode("utf8")) + tmp2.write(("".join(submitseq)).encode("utf8")) + tmp2.flush() + subbp = 0 + refbp = 0 + similarity = 0 + try: + cmd = ["minimap2", "-c", tmp1.name, tmp2.name] + logging.info("QC checking similarity to reference") + logging.info(" ".join(cmd)) + result = subprocess.run(cmd, stdout=subprocess.PIPE) + result.check_returncode() + res = result.stdout.decode("utf-8") + mm = res.split("\t") + if len(mm) >= 10: + # divide Number of matching bases in the mapping / Target sequence length + similarity = (float(mm[9]) / float(mm[6])) * 100.0 + else: + similarity = 0 + except Exception as e: + logging.warn("QC against reference sequence using 'minimap2': %s", e, exc_info=e) - if refbp and (subbp/refbp) < .7: - raise ValueError("QC fail: submit sequence length is shorter than 70% reference") - if refbp and (subbp/refbp) > 1.3: - raise ValueError("QC fail: submit sequence length is greater than 130% reference") - if similarity and similarity < 70.0: - raise ValueError("QC fail: submit similarity is less than 70%") - if refbp == 0 or similarity == 0: - raise ValueError("QC fail") + if similarity and similarity < 70.0: + raise ValueError("QC fail: alignment to reference was less than 70%% (was %2.2f%%)" % (similarity)) + if similarity == 0: + raise ValueError("QC fail") return ("sequence.fasta"+gz, seqlabel) elif seq_type == "text/fastq": + sequence.seek(0) + sequence.detach() return ("reads.fastq"+gz, seqlabel) else: raise ValueError("Sequence file does not look like a DNA FASTA or FASTQ") |