mgnify-pipelines-toolkit 1.2.0__py3-none-any.whl → 1.2.1__py3-none-any.whl

mgnify_pipelines_toolkit/analysis/amplicon/standard_primer_matching.py

@@ -1,327 +0,0 @@
-#!/usr/bin/env python
-# -*- coding: utf-8 -*-
-
-# Copyright 2024-2025 EMBL - European Bioinformatics Institute
-#
-# Licensed under the Apache License, Version 2.0 (the "License");
-# you may not use this file except in compliance with the License.
-# You may obtain a copy of the License at
-# http://www.apache.org/licenses/LICENSE-2.0
-#
-# Unless required by applicable law or agreed to in writing, software
-# distributed under the License is distributed on an "AS IS" BASIS,
-# WITHOUT WARRANTIES OR CONDITIONS OF ANY KIND, either express or implied.
-# See the License for the specific language governing permissions and
-# limitations under the License.
-
-import argparse
-from collections import defaultdict
-import os
-
-from Bio.Seq import Seq
-import regex
-
-from mgnify_pipelines_toolkit.analysis.amplicon.amplicon_utils import (
-    primer_regex_query_builder,
-    get_read_count,
-    fetch_mcp,
-)
-
-
-def parse_args():
-
-    parser = argparse.ArgumentParser()
-
-    parser.add_argument(
-        "-i",
-        "--input",
-        required=True,
-        type=str,
-        help="Path to merged FASTQ to look for primers",
-    )
-    parser.add_argument(
-        "-p",
-        "--primers",
-        required=True,
-        type=str,
-        help="Path to directory containing standard primers fasta files",
-    )
-    parser.add_argument("-s", "--sample", required=True, type=str, help="Sample ID")
-    parser.add_argument("-o", "--output", required=True, type=str, help="Output path")
-    args = parser.parse_args()
-
-    input = args.input
-    primers = args.primers
-    sample = args.sample
-    output = args.output
-
-    return input, primers, sample, output
-
-
-def parse_std_primers(primers):
-    """
-    Parse the library of standard primers.
-
-    Reads the fasta files in the given directory
-    Primer names (which are the fasta headers) are labeled with F or R for 5'-3' and 3'-5' primers respectively
-
-    Returns two dictionaries:
-        std_primer_dict_regex
-            key: region+primer name
-            val: primer sequence from 5' to 3'
-        std_primer_dict
-            key: region+primer name
-            val: primer sequence from 5' to 3' for forward primers, 3' to 5' for reverse
-    """
-
-    std_primer_dict_regex = defaultdict(defaultdict)
-    std_primer_dict = defaultdict(defaultdict)
-
-    dir = os.listdir(primers)
-    dir = [f"{primers}/{path}" for path in dir]
-
-    rev_flag = False
-
-    for path in dir:
-        region = path.split("/")[-1].split(".")[0]
-        with open(path, "r") as fr:
-            key = ""
-            for line in fr:
-                line = line.strip()
-                if line[0] == ">":
-                    if "R" in line:  # If a primer is a reverse primer
-                        rev_flag = True
-                    key = line[1:]
-                else:
-                    if rev_flag:
-                        rev_conv = str(Seq(line).complement())
-                        line = rev_conv
-                        rev_flag = False
-
-                    primer = primer_regex_query_builder(line)
-                    std_primer_dict_regex[region][key] = primer
-                    std_primer_dict[region][key] = line
-
-    return std_primer_dict_regex, std_primer_dict
-
-
-def run_primer_matching_once(input_path, input_primer, rev=False):
-    """
-    Run primer matching using the regex package.
-
-    Takes one primer, strand, and fastq input
-    Uses fuzzy matching to allow for at most one error (for sequencing errors)
-    Returns number of reads matching given primer
-    """
-
-    match_count = 0.0
-
-    mcp_count_dict = fetch_mcp(input_path, 50, rev=rev)
-
-    for mcp in mcp_count_dict.keys():
-        mcp = mcp.strip()
-        res = regex.match(input_primer, mcp)
-        if res is not None:
-            match_count += mcp_count_dict[mcp]
-
-    return match_count
-
-
-def get_primer_props(std_primer_dict_regex, input_path):
-    """
-    Look for the standard primers in the input fastq file.
-
-    Will loop through the dictionary of primers, using fuzzy regex matching to find matching primers.
-    If a std primer is present above a set threshold proportion, it is collected. Both strands are searched for.
-    If there is an std primer for both the F and R strands, the maximum prop for each strand is chosen and the pair
-    is output as a combination.
-
-    Returns a list containing two elements:
-        max_region: the amplified region the chosen primers belong to
-        max_primers: dictionary containing the F and/or R primers that were chosen
-    """
-
-    threshold = 0.60  # Arbitrary threshold for collecting a matched primer
-    read_count = get_read_count(
-        input_path, file_type="fastq"
-    )  # Get read count of fastq file to calculate proportion with
-    res_dict = defaultdict(defaultdict)
-
-    # Loop through every primer region
-    for region, primer in std_primer_dict_regex.items():
-        res_dict[region]["F"] = {}
-        res_dict[region]["R"] = {}
-
-        # Loop through every primer of a certain region
-        for primer_name, primer_seq in primer.items():
-
-            region_name_str = f"{region};{primer_name}"
-            primer_count = 0.0
-
-            if "F" in primer_name:
-                primer_count = run_primer_matching_once(
-                    input_path, primer_seq, rev=False
-                )  # Get proportion of a F primer with fuzzy regex matching
-            elif "R" in primer_name:
-                primer_count = run_primer_matching_once(
-                    input_path, primer_seq, rev=True
-                )  # Get proportion of a R primer with fuzzy regex matching
-
-            try:
-                primer_prop = primer_count / read_count
-            except ZeroDivisionError:
-                primer_prop = 0
-
-            if "F" in primer_name:
-                if (
-                    primer_prop > threshold
-                ):  # Only collect primer if it's above threshold
-                    res_dict[region]["F"][primer_name] = primer_prop
-            elif "R" in primer_name:
-                if (
-                    primer_prop > threshold
-                ):  # Only collect primer if it's above threshold
-                    res_dict[region]["R"][primer_name] = primer_prop
-
-            print(f"{region_name_str}: {primer_prop}")
-
-        # If an F or/and R primer wasn't found then just remove it from the dictionary
-        if res_dict[region]["F"] == {}:
-            res_dict[region].pop("F")
-        if res_dict[region]["R"] == {}:
-            res_dict[region].pop("R")
-
-    singles = defaultdict(str)
-    doubles = defaultdict(list)
-
-    double_status = False  # Flag for whether primers were found on both strands
-
-    #  Loop through every collected primer and put primers in singles or doubles
-    for region in res_dict.keys():
-        strands = res_dict[region]
-
-        for strand in strands.keys():
-            primers = strands[strand]
-            max_prop = 0
-            max_name = ""
-            for primer_name, prop in primers.items():
-                if prop > max_prop:
-                    max_prop = prop
-                    max_name = primer_name
-
-            if len(strands.keys()) == 2:
-                double_status = True
-                doubles[region].append({max_name: max_prop})
-            elif len(strands.keys()) == 1:
-                singles[region] = {max_name: max_prop}
-
-    max_region = ""
-    max_primers = {}
-    max_mean_prop = 0
-
-    # if at least one pair of primers was collected
-    if double_status:
-        for (
-            region
-        ) in doubles:  # Loop through all pairs of primers and choose the best one
-            primers = doubles[region]
-
-            f_primer_name = list(primers[0].keys())[0]
-            r_primer_name = list(primers[1].keys())[0]
-            f_primer_prop = primers[0][f_primer_name]
-            r_primer_prop = primers[1][r_primer_name]
-
-            mean_prop = (f_primer_prop + r_primer_prop) / 2.0
-            if mean_prop > max_mean_prop:
-                max_mean_prop = mean_prop
-                max_region = region
-                max_primers = [
-                    {f_primer_name: f_primer_prop},
-                    {r_primer_name: r_primer_prop},
-                ]
-
-    else:
-        for region in singles:  # Choose the best single primer
-            primer = singles[region]
-            primer_name = list(primer.keys())[0]
-            prop = primer[primer_name]
-            if prop > max_mean_prop:
-                max_mean_prop = prop
-                max_region = region
-                max_primers = {primer_name: prop}
-
-    if max_region == "":
-        print("No standard library primers!")
-        return []
-    elif double_status:
-        print("Standard library primers found!")
-        print(f"Region: {max_region}")
-        print(f"Forward Primer: {max_primers[0]}")
-        print(f"Reverse Primer: {max_primers[1]}")
-
-        return [max_region, max_primers[0], max_primers[1]]
-    else:
-        print("Standard library primer found on one strand!")
-        print(f"Region: {max_region}")
-        print(f"Primer: {max_primers}")
-
-        return [max_region, max_primers]
-
-
-def save_out(results, sample_id, output, std_primer_dict):
-    """
-    Save found std primers into a fasta file.
-    """
-
-    with (
-        open(f"{output}/{sample_id}_std_primer_out.txt", "w") as fw_out,
-        open(f"{output}/{sample_id}_std_primers.fasta", "w") as fw_seq,
-    ):
-        if results == []:
-            fw_out.write("")
-            fw_seq.write("")
-
-        elif len(results) == 2:
-            region = results[0]
-            primer_name = list(results[1].keys())[0]
-            primer_prop = results[1][list(results[1].keys())[0]]
-            seq = std_primer_dict[region][primer_name]
-            if "R" in primer_name:
-                seq = str(Seq(seq).complement())
-            fw_out.write(f"{region}\n")
-            fw_out.write(f"{primer_name}: {primer_prop}")
-
-            fw_seq.write(f">{primer_name}\n{seq}")
-
-        elif len(results) == 3:
-            region = results[0]
-            f_primer_name = list(results[1].keys())[0]
-            f_primer_prop = results[1][list(results[1].keys())[0]]
-            f_seq = std_primer_dict[region][f_primer_name]
-            r_primer_name = list(results[2].keys())[0]
-            r_primer_prop = results[2][list(results[2].keys())[0]]
-            r_seq = std_primer_dict[region][r_primer_name]
-            r_seq = str(Seq(r_seq).complement())
-
-            fw_out.write(f"{region}\n")
-            fw_out.write(f"{f_primer_name}: {f_primer_prop}\n")
-            fw_out.write(f"{r_primer_name}: {r_primer_prop}")
-
-            fw_seq.write(f">{f_primer_name}\n{f_seq}\n")
-            fw_seq.write(f">{r_primer_name}\n{r_seq}\n")
-
-
-def main():
-
-    input, primers, sample, output = parse_args()
-    std_primer_dict_regex, std_primer_dict = parse_std_primers(
-        primers
-    )  # Parse std primer library into dictionaries
-    results = get_primer_props(
-        std_primer_dict_regex, input
-    )  # Find all the std primers in the input and select most common
-    save_out(results, sample, output, std_primer_dict)
-
-
-if __name__ == "__main__":
-    main()

mgnify_pipelines_toolkit/constants/regex_ambiguous_bases.py

@@ -1,43 +0,0 @@
-#!/usr/bin/env python
-# -*- coding: utf-8 -*-
-
-# Copyright 2024-2025 EMBL - European Bioinformatics Institute
-#
-# Licensed under the Apache License, Version 2.0 (the "License");
-# you may not use this file except in compliance with the License.
-# You may obtain a copy of the License at
-# http://www.apache.org/licenses/LICENSE-2.0
-#
-# Unless required by applicable law or agreed to in writing, software
-# distributed under the License is distributed on an "AS IS" BASIS,
-# WITHOUT WARRANTIES OR CONDITIONS OF ANY KIND, either express or implied.
-# See the License for the specific language governing permissions and
-# limitations under the License.
-
-_AMBIGUOUS_BASES_DICT = {
-    "R": "[AG]",
-    "Y": "[CT]",
-    "S": "[GC]",
-    "W": "[AT]",
-    "K": "[GT]",
-    "M": "[AC]",
-    "B": "[CGT]",
-    "D": "[AGT]",
-    "H": "[ACT]",
-    "V": "[ACG]",
-    "N": "[ACTG]",
-}
-
-_AMBIGUOUS_BASES_DICT_REV = {
-    "A,G": "R",
-    "C,T": "Y",
-    "C,G": "S",
-    "A,T": "W",
-    "G,T": "K",
-    "A,C": "M",
-    "C,G,T": "B",
-    "A,G,T": "D",
-    "A,C,T": "H",
-    "A,C,G": "V",
-    "A,C,G,T": "N",
-}