Completed NCBI lookup, FASTA reading, ABIF reading, and Institut Pasteur querying

requirements.txt

@@ -2,3 +2,4 @@ aiohttp[speedups]
 biopython
 pytest
 pytest-asyncio
+dataclasses-json

src/mlstmyfasta/engine/annotate.py

@@ -6,30 +6,8 @@ from Bio import Entrez
 from Bio import SeqIO
 import numpy as np
 
-
-# TODO Change this out for a more professional approach
-Entrez.email = "yunyangdeng@outlook.com"
-
-from mlstmyfasta.engine.data.genomics import AnnotatedString, StringAnnotation, get_feature_coding
-
-
-async def fetch_ncbi_genbank(genbank_id: str) -> AnnotatedString:
-    with (await asyncio.to_thread(Entrez.efetch, db="nucleotide", id=genbank_id, rettype="gb", retmode="text")) as fetch_stream:
-        record = SeqIO.read(fetch_stream, "genbank")
-        sequence_features = list()
-        for feature in record.features:
-            start = int(feature.location.start)
-            end = int(feature.location.end)
-            qualifiers = feature.qualifiers
-            for qualifier_key in qualifiers:
-                qualifiers[qualifier_key] = set(qualifiers[qualifier_key])
-            sequence_features.append(StringAnnotation(
-                type=feature.type,
-                start=start,
-                end=end+1,  # Position is exclusive
-                feature_properties=qualifiers
-            ))
-        return AnnotatedString(name=genbank_id, sequence=str(record.seq), annotations=sequence_features)
+from mlstmyfasta.engine.data.genomics import AnnotatedString, StringAnnotation
+from mlstmyfasta.engine.remote.databases.ncbi.genbank import fetch_ncbi_genbank
 
 
 async def annotate_from_genbank(genbank_id: str, query_name: str, query_string: str, max_annotation_length:int = 512, gene_targets:Set = set()):
@@ -61,3 +39,6 @@ async def annotate_from_genbank(genbank_id: str, query_name: str, query_string:
             feature_properties=dict(annotation.feature_properties) # same as original
         ))
     return AnnotatedString(name=query_name, sequence=query_string, annotations=query_annotations)
+
+def get_feature_coding(annotated_string: AnnotatedString, string_annotation: StringAnnotation) -> str:
+    return annotated_string.sequence[string_annotation.start:string_annotation.end]

src/mlstmyfasta/engine/data/MLST.py

@@ -2,5 +2,5 @@ from dataclasses import dataclass
 
 @dataclass
 class Allele:
-    allele_name: str
+    allele_loci: str
     allele_variant: str

src/mlstmyfasta/engine/data/genomics.py

@@ -1,5 +1,6 @@
 from dataclasses import dataclass
-from typing import Mapping, Sequence, Set
+from numbers import Number
+from typing import Mapping, Sequence, Set, Union
 
 
 @dataclass
@@ -10,10 +11,95 @@ class StringAnnotation:
     feature_properties: Mapping[str, Set[str]]
 
 @dataclass
-class AnnotatedString:
+class NamedString:
     name: str
     sequence: str
+
+@dataclass
+class AnnotatedString(NamedString):
     annotations: Sequence[StringAnnotation]
 
-def get_feature_coding(annotated_string: AnnotatedString, string_annotation: StringAnnotation) -> str:
-    return annotated_string.sequence[string_annotation.start:string_annotation.end]
+@dataclass
+class SangerTraceData:
+    sequence: Sequence[str]
+    seq_param_file_name: str
+    analysis_proto_settings_name: str
+    analysis_rpto_settings_ver: str
+    analysis_proto_xml_data: str
+    analysis_proto_xml_schema_ver: str
+    sample_comment: Union[None, str]
+    capillary_machine: bool
+    container_identifier: str
+    container_name: str
+    comment_title: str
+    channel_1: Sequence[Number]
+    channel_2: Sequence[Number]
+    channel_3: Sequence[Number]
+    channel_4: Sequence[Number]
+    measured_voltage_dv: Sequence[Number]
+    measured_current_ma: Sequence[Number]
+    measured_power_mw: Sequence[Number]
+    measured_temperature_celsius: Sequence[Number]
+    down_sample_factor: Number
+    dye_1: str
+    dye_2: str
+    dye_3: str
+    dye_4: str
+    dye_wavelength_1: str
+    dye_wavelength_2: str
+    dye_wavelength_3: str
+    dye_wavelength_4: str
+    dye_set_name: str
+    electrophoresis_voltage_setting_v: Number
+    start_run_event: str
+    stop_run_event: str
+    start_collection_event: str
+    stop_collection_event: str
+    base_order: Sequence[str]
+    gel_type_desc: str
+    injection_time_sec: Number
+    inection_voltage_v: Number
+    lane_or_capillary: Number
+    sample_tracking_id: str
+    length_to_detector_cm: Number
+    laser_power_mw: Number
+    instrument_name_and_serial: str
+    data_collection_module_file: str
+    model_number: str
+    pixels_avg_per_lane: Number
+    number_of_capillaries: Number
+    marked_off_scale_scans: Union[None, Sequence[Number]]
+    # Skipped Ovrl, OvrV
+    mobility_file: str
+    # Skipped PRJT, PROJ
+    pixel_bin_size: Number
+    # Skipped scan rate
+    results_group_comment: Union[None, str]
+    results_group_name: str
+    run_module_ver: str
+    run_module_xml: str
+    run_module_xml_ver: str
+    run_proto_name: str
+    run_proto_ver: str
+    run_start_date: str  # Date time object
+    run_stop_date: str  # Date time object
+    data_collection_start_date: str
+    data_collection_stop_date: str
+    run_name: str
+    run_start_time: str  # time object
+    run_stop_time: str  # time object
+    collection_start_time: str  # time object
+    collection_stop_time: str  # time object
+    saturated_data_points: Union[None, Sequence[Number]]
+    color_rescaling_divisor: Number
+    scan_count: Number
+    polymer_lot_expiration: str  # date time object
+    polymer_lot_number: Number
+    sample_name: str
+    # Skipped genescan data
+    # Skipped size standard file name
+    data_collection_software_ver: str
+    data_collection_firmware_ver: str
+    run_temperature_setting_celcius: Number
+    well_id: str
+    plate_user_name: str

src/mlstmyfasta/engine/local/abif.py

@@ -0,0 +1,104 @@
+import asyncio
+from numbers import Number
+from os import path
+from typing import Sequence, Union
+from mlstmyfasta.engine.data.genomics import SangerTraceData
+from Bio.SeqRecord import SeqRecord
+from Bio import SeqIO
+
+
+def _biopython_read_abif_sequence(seq_path: str) -> SeqRecord:
+    with open(seq_path, "rb") as seq_handle:
+        return SeqIO.read(seq_handle, "abi")
+
+
+async def load_sanger_sequence(seq_path: str) -> SangerTraceData:
+    ext = path.splitext(seq_path)[1]
+    if ext.lower() != ".ab1" and ext.lower() != "abi":
+        raise ValueError(
+            'seq_path must have file extension of "ab1", or "abi".')
+    biopython_seq = await asyncio.to_thread(_biopython_read_abif_sequence, seq_path)
+    biopython_annotations = biopython_seq.annotations
+
+    # Lot of type ignoring since Biopython did not define their typing.
+    biopython_abif_raw = biopython_annotations["abif_raw"] # type: ignore
+    trace_data = SangerTraceData(
+        biopython_seq.seq,
+        biopython_abif_raw.get("APFN2"), # type: ignore
+        biopython_abif_raw.get("APrN1"), # type: ignore
+        biopython_abif_raw.get("APrV1"), # type: ignore
+        biopython_abif_raw.get("APrX1"), # type: ignore
+        biopython_abif_raw.get("APXV1"), # type: ignore
+        biopython_abif_raw.get("CMNT1"), # type: ignore
+        biopython_abif_raw.get("CpEP1"), # type: ignore
+        biopython_abif_raw.get("CTID1"), # type: ignore
+        biopython_abif_raw.get("CTNM1"), # type: ignore
+        biopython_abif_raw.get("CTTL1"), # type: ignore
+        biopython_abif_raw.get("DATA1"), # type: ignore
+        biopython_abif_raw.get("DATA2"), # type: ignore
+        biopython_abif_raw.get("DATA3"), # type: ignore
+        biopython_abif_raw.get("DATA4"), # type: ignore
+        biopython_abif_raw.get("DATA5"), # type: ignore
+        biopython_abif_raw.get("DATA6"), # type: ignore
+        biopython_abif_raw.get("DATA7"), # type: ignore
+        biopython_abif_raw.get("DATA8"), # type: ignore
+        biopython_abif_raw.get("DSam1"), # type: ignore
+        biopython_abif_raw.get("DyeN1"), # type: ignore
+        biopython_abif_raw.get("DyeN2"), # type: ignore
+        biopython_abif_raw.get("DyeN3"), # type: ignore
+        biopython_abif_raw.get("DyeN4"), # type: ignore
+        biopython_abif_raw.get("DyeW1"), # type: ignore
+        biopython_abif_raw.get("DyeW2"), # type: ignore
+        biopython_abif_raw.get("DyeW3"), # type: ignore
+        biopython_abif_raw.get("DyeW4"), # type: ignore
+        biopython_abif_raw.get("DySN1"), # type: ignore
+        biopython_abif_raw.get("EPVt1"), # type: ignore
+        biopython_abif_raw.get("EVNT1"), # type: ignore
+        biopython_abif_raw.get("EVNT2"), # type: ignore
+        biopython_abif_raw.get("EVNT3"), # type: ignore
+        biopython_abif_raw.get("EVNT4"), # type: ignore
+        biopython_abif_raw.get("FWO_1"), # type: ignore
+        biopython_abif_raw.get("GTyp1"), # type: ignore
+        biopython_abif_raw.get("InSc1"), # type: ignore
+        biopython_abif_raw.get("InVt1"), # type: ignore
+        biopython_abif_raw.get("LANE1"), # type: ignore
+        biopython_abif_raw.get("LIMS1"), # type: ignore
+        biopython_abif_raw.get("LNTD1"), # type: ignore
+        biopython_abif_raw.get("LsrP1"), # type: ignore
+        biopython_abif_raw.get("MCHN1"), # type: ignore
+        biopython_abif_raw.get("MODF1"), # type: ignore
+        biopython_abif_raw.get("MODL1"), # type: ignore
+        biopython_abif_raw.get("NAVG1"), # type: ignore
+        biopython_abif_raw.get("NLNE1"), # type: ignore
+        biopython_abif_raw.get("OfSc1"), # type: ignore
+        biopython_abif_raw.get("PDMF1"), # type: ignore
+        biopython_abif_raw.get("PXLB1"), # type: ignore
+        biopython_abif_raw.get("RGCm1"), # type: ignore
+        biopython_abif_raw.get("RGNm1"), # type: ignore
+        biopython_abif_raw.get("RMdV1"), # type: ignore
+        biopython_abif_raw.get("RMdX1"), # type: ignore
+        biopython_abif_raw.get("RMXV1"), # type: ignore
+        biopython_abif_raw.get("RPrN1"), # type: ignore
+        biopython_abif_raw.get("RPrV1"), # type: ignore
+        biopython_abif_raw.get("RUND1"), # type: ignore
+        biopython_abif_raw.get("RUND2"), # type: ignore
+        biopython_abif_raw.get("RUND3"), # type: ignore
+        biopython_abif_raw.get("RUND4"), # type: ignore
+        biopython_abif_raw.get("RunN1"), # type: ignore
+        biopython_abif_raw.get("RUNT1"), # type: ignore
+        biopython_abif_raw.get("RUNT2"), # type: ignore
+        biopython_abif_raw.get("RUNT3"), # type: ignore
+        biopython_abif_raw.get("RUNT4"), # type: ignore
+        biopython_abif_raw.get("Satd"), # type: ignore
+        biopython_abif_raw.get("Scal1"), # type: ignore
+        biopython_abif_raw.get("SCAN1"), # type: ignore
+        biopython_abif_raw.get("SMED1"), # type: ignore
+        biopython_abif_raw.get("SMLt"), # type: ignore
+        biopython_abif_raw.get("SMPL1"), # type: ignore
+        biopython_abif_raw.get("SVER1"), # type: ignore
+        biopython_abif_raw.get("SVER3"), # type: ignore
+        biopython_abif_raw.get("Tmpr1"), # type: ignore
+        biopython_abif_raw.get("TUBE"), # type: ignore
+        biopython_abif_raw.get("User") # type: ignore
+    )
+    return trace_data

src/mlstmyfasta/engine/local/fasta.py

@@ -0,0 +1,11 @@
+import asyncio
+from io import TextIOWrapper
+from typing import Any, AsyncGenerator, Generator, Sequence, Union
+from Bio import SeqIO
+
+from mlstmyfasta.engine.data.genomics import NamedString
+
+async def read_fasta(handle: Union[str, TextIOWrapper]) -> AsyncGenerator[NamedString, Any]:
+    fasta_sequences = asyncio.to_thread(SeqIO.parse, handle=handle, format="fasta")
+    for fasta_sequence in await fasta_sequences:
+        yield NamedString(fasta_sequence.id, str(fasta_sequence.seq))

src/mlstmyfasta/engine/remote/databases/institutpasteur/profiling.py

@@ -0,0 +1,36 @@
+from contextlib import AbstractAsyncContextManager
+import re
+from typing import Sequence
+from aiohttp import ClientSession, ClientTimeout
+from mlstmyfasta.engine.data.MLST import Allele
+from mlstmyfasta.engine.data.genomics import NamedString
+from mlstmyfasta.engine.remote.databases.institutpasteur.structures import InstitutPasteurSequenceRequest, InstitutPasteurSequenceResponse
+
+class InstitutPasteurProfiler(AbstractAsyncContextManager):
+
+    async def __aenter__(self):
+        return self
+
+
+    def __init__(self, database_name: str):
+        self._base_url = f"https://bigsdb.pasteur.fr/api/db/{database_name}/"
+        self._http_client = ClientSession(self._base_url, timeout=ClientTimeout(10000))
+
+    async def fetch_mlst_profile(self, sequence_string: str):
+        # See https://bigsdb.pasteur.fr/api/db/pubmlst_bordetella_seqdef/schemes
+        uri_path = f"schemes/3/sequence"
+        response = await self._http_client.post(uri_path, json={
+            "sequence": sequence_string
+        })
+        sequence_response: dict = await response.json()
+        exact_matches: dict[str, Sequence[dict[str, str]]] = sequence_response["exact_matches"]  
+        for allele_loci, alleles in exact_matches.items():
+            for allele in alleles:
+                alelle_id = allele["allele_id"]
+                yield Allele(allele_loci=allele_loci, allele_variant=alelle_id)
+
+    async def close(self):
+        await self._http_client.close()
+
+    async def __aexit__(self, exc_type, exc_value, traceback):
+        await self.close()

src/mlstmyfasta/engine/remote/databases/institutpasteur/structures.py

@@ -0,0 +1,22 @@
+from dataclasses import dataclass
+from typing import Sequence, Union
+
+from dataclasses_json import dataclass_json
+
+@dataclass_json
+@dataclass
+class InstitutPasteurSequenceRequest:
+    sequence: str
+    details: Union[bool, None]
+    partial_matches: Union[bool, None]
+    base64: Union[bool, None]
+
+@dataclass_json
+@dataclass
+class InstitutPasteurSequenceResponse:
+    exact_matches: Sequence[tuple[int, str]]
+    start: Union[int, None]
+    end: Union[int, None]
+    orientation: Union[str, None]
+    length: Union[int, None]
+    contig: Union[str, None]

src/mlstmyfasta/engine/remote/databases/ncbi/genbank.py

@@ -0,0 +1,27 @@
+import asyncio
+from Bio import Entrez
+from Bio import SeqIO
+
+# TODO Change this out for a more professional approach
+Entrez.email = "yunyangdeng@outlook.com"
+
+from mlstmyfasta.engine.data.genomics import AnnotatedString, StringAnnotation
+
+
+async def fetch_ncbi_genbank(genbank_id: str) -> AnnotatedString:
+    with (await asyncio.to_thread(Entrez.efetch, db="nucleotide", id=genbank_id, rettype="gb", retmode="text")) as fetch_stream:
+        record = SeqIO.read(fetch_stream, "genbank")
+        sequence_features = list()
+        for feature in record.features:
+            start = int(feature.location.start)
+            end = int(feature.location.end)
+            qualifiers = feature.qualifiers
+            for qualifier_key in qualifiers:
+                qualifiers[qualifier_key] = set(qualifiers[qualifier_key])
+            sequence_features.append(StringAnnotation(
+                type=feature.type,
+                start=start,
+                end=end+1,  # Position is exclusive
+                feature_properties=qualifiers
+            ))
+        return AnnotatedString(name=genbank_id, sequence=str(record.seq), annotations=sequence_features)

src/mlstmyfasta/engine/web/databases/institutpasteur/structures.py

@@ -1,18 +0,0 @@
-from dataclasses import dataclass
-from typing import Sequence, Union
-
-@dataclass
-class SequenceRequestBody:
-    sequence: str
-    details: Union[None, str]
-    partial_matches: Union[None, bool]
-    base64: Union[None, bool]
-
-@dataclass
-class SequenceResponse:
-    exact_matches: Sequence[tuple[str, str]]
-    start: Union[None, int]
-    end: Union[None, int]
-    orientation: Union[None, str]
-    length: Union[None, int]
-    contig: Union[None, str]

tests/mlstmyfasta/engine/local/test_abif.py

@@ -0,0 +1,8 @@
+import os
+
+from mlstmyfasta.engine.local.abif import load_sanger_sequence
+
+async def test_load_sanger_sequence_has_data():
+    assert os.path.exists("tests/resources/1I1_F_P1815443_047.ab1")
+    result_data = await load_sanger_sequence("tests/resources/1I1_F_P1815443_047.ab1")
+    assert result_data is not None

tests/mlstmyfasta/engine/local/test_fasta.py

@@ -0,0 +1,7 @@
+from mlstmyfasta.engine.local.fasta import read_fasta
+
+
+async def test_fasta_reader_not_none():
+    named_strings = read_fasta("tests/resources/tohama_I_bpertussis.fasta")
+    async for named_string in named_strings:
+        assert named_string.name == "BX470248.1"

tests/mlstmyfasta/engine/remote/databases/institutpasteur/test_profiles.py

@@ -0,0 +1,16 @@
+from Bio import SeqIO
+from mlstmyfasta.engine.data.MLST import Allele
+from mlstmyfasta.engine.remote.databases.institutpasteur.profiling import InstitutPasteurProfiler
+
+
+async def test_profiling_results_in_exact_matches_when_exact():
+    sequence = str(SeqIO.read("tests/resources/tohama_I_bpertussis.fasta", "fasta").seq)
+    async with InstitutPasteurProfiler(database_name="pubmlst_bordetella_seqdef") as dummy_profiler:
+        exact_matches = dummy_profiler.fetch_mlst_profile(sequence_string=sequence)
+        targets_left = {"adk", "fumC", "glyA", "tyrB", "icd", "pepA", "pgm"}
+        async for exact_match in exact_matches:
+            assert isinstance(exact_match, Allele)
+            assert exact_match.allele_variant == '1' # All of Tohama I has allele id I
+            targets_left.remove(exact_match.allele_loci)
+
+        assert len(targets_left) == 0

tests/mlstmyfasta/engine/remote/databases/ncbi/test_genbank.py

@@ -0,0 +1,5 @@
+from mlstmyfasta.engine.remote.databases.ncbi.genbank import fetch_ncbi_genbank
+
+
+async def test_fetch_ncbi_genbank_with_id_works():
+    assert len((await fetch_ncbi_genbank("CP011448.1")).sequence) > 0 

tests/mlstmyfasta/engine/test_annotate.py

@@ -1,14 +1,11 @@
-from mlstmyfasta.engine.annotations import annotate_from_genbank, fetch_ncbi_genbank
+from mlstmyfasta.engine.annotate import annotate_from_genbank, fetch_ncbi_genbank
 from Bio import SeqIO
 
 from mlstmyfasta.engine.data.genomics import AnnotatedString
 
-async def test_fetch_ncbi_genbank_with_id_works():
-    assert len((await fetch_ncbi_genbank("CP011448.1")).sequence) > 0 
-
 async def test_annotate_from_genbank_for_adk_annotation():
-    sequence = SeqIO.read("tests/resources/tohama_I_bpertussis.fasta", "fasta").seq
-    annotated_sequence = await annotate_from_genbank("CP011448.1", "bpertussis_tohamaI", str(sequence), max_annotation_length=750, gene_targets=set(["adk"]))
+    sequence = str(SeqIO.read("tests/resources/tohama_I_bpertussis.fasta", "fasta").seq)
+    annotated_sequence = await annotate_from_genbank("CP011448.1", "bpertussis_tohamaI", sequence, max_annotation_length=750, gene_targets=set(["adk"]))
     assert isinstance(annotated_sequence, AnnotatedString)
     assert len(annotated_sequence.annotations) >= 1
     assert annotated_sequence.annotations[0].type == "gene"