From 981057dfda08c33d42e0753a817dc823c5dc714f Mon Sep 17 00:00:00 2001 From: Nils Homer Date: Wed, 11 Dec 2024 12:35:05 -0700 Subject: [PATCH] feat: initial implementation of python bindings for `bwa aln` --- .github/CODEOWNERS | 1 + .github/workflows/ci.yml | 90 + .github/workflows/readthedocs.yaml | 20 + .readthedocs.yml | 12 + README.md | 28 + build.py | 112 + bwapy.yml | 9 + bwapy/__init__.py | 4 + bwapy/libbwapy.pxd | 119 + bwapy/libbwapy.pyi | 42 + bwapy/libbwapy.pyx | 356 + bwapy/libbwapy_utils.c | 25 + bwapy/libbwapy_utils.h | 8 + bwapy/py.typed | 0 codecov.yml | 2 + docs/index.md | 9 + ...stallation-and-developers-documentation.md | 76 + docs/scripts/gen_ref_pages.py | 49 + mkdocs.yml | 52 + poetry.lock | 1648 + pyproject.toml | 94 + tests/data/e_coli_k12.dict | 2 + tests/data/e_coli_k12.fasta | 57997 ++++++++++++++++ tests/data/e_coli_k12.fasta.amb | 1 + tests/data/e_coli_k12.fasta.ann | 3 + tests/data/e_coli_k12.fasta.bwt | Bin 0 -> 4639752 bytes tests/data/e_coli_k12.fasta.fai | 1 + tests/data/e_coli_k12.fasta.pac | Bin 0 -> 1159920 bytes tests/data/e_coli_k12.fasta.sa | Bin 0 -> 2319888 bytes tests/test_bwapy.py | 44 + 30 files changed, 60804 insertions(+) create mode 100644 .github/CODEOWNERS create mode 100644 .github/workflows/ci.yml create mode 100644 .github/workflows/readthedocs.yaml create mode 100644 .readthedocs.yml create mode 100644 build.py create mode 100644 bwapy.yml create mode 100644 bwapy/__init__.py create mode 100644 bwapy/libbwapy.pxd create mode 100644 bwapy/libbwapy.pyi create mode 100644 bwapy/libbwapy.pyx create mode 100644 bwapy/libbwapy_utils.c create mode 100644 bwapy/libbwapy_utils.h create mode 100644 bwapy/py.typed create mode 100644 codecov.yml create mode 100644 docs/index.md create mode 100644 docs/installation-and-developers-documentation.md create mode 100644 docs/scripts/gen_ref_pages.py create mode 100644 mkdocs.yml create mode 100644 poetry.lock create mode 100644 pyproject.toml create mode 100755 tests/data/e_coli_k12.dict create mode 100755 tests/data/e_coli_k12.fasta create mode 100755 tests/data/e_coli_k12.fasta.amb create mode 100755 tests/data/e_coli_k12.fasta.ann create mode 100755 tests/data/e_coli_k12.fasta.bwt create mode 100755 tests/data/e_coli_k12.fasta.fai create mode 100755 tests/data/e_coli_k12.fasta.pac create mode 100755 tests/data/e_coli_k12.fasta.sa create mode 100644 tests/test_bwapy.py diff --git a/.github/CODEOWNERS b/.github/CODEOWNERS new file mode 100644 index 0000000..77092ae --- /dev/null +++ b/.github/CODEOWNERS @@ -0,0 +1 @@ +* @nh13 diff --git a/.github/workflows/ci.yml b/.github/workflows/ci.yml new file mode 100644 index 0000000..0e8fdf3 --- /dev/null +++ b/.github/workflows/ci.yml @@ -0,0 +1,90 @@ +name: CI + +on: push +env: + POETRY_VERSION: 1.8 + +jobs: + testing: + runs-on: ubuntu-24.04 + strategy: + matrix: + PYTHON_VERSION: ["3.9", "3.10", "3.11", "3.12"] + steps: + - uses: actions/checkout@v4 + - uses: actions/checkout@v4 + with: + repository: "lh3/bwa" + path: "bwa" + - name: Set up Python ${{matrix.PYTHON_VERSION}} + uses: actions/setup-python@v1 + with: + python-version: ${{matrix.PYTHON_VERSION}} + + - name: Get full Python version + id: full-python-version + shell: bash + run: echo ::set-output name=version::$(python -c "import sys; print('-'.join(str(v) for v in sys.version_info))") + + - name: Install poetry + shell: bash + run: | + python -m pip install --upgrade pip + pip install poetry==${{env.POETRY_VERSION}} + + - name: Configure poetry + shell: bash + run: poetry config virtualenvs.in-project true + + - name: Set up cache + uses: actions/cache@v2 + id: cache + with: + path: .venv + key: venv-${{ runner.os }}-${{ steps.full-python-version.outputs.version }}-${{ hashFiles('**/poetry.lock') }} + + - name: Ensure cache is healthy + if: steps.cache.outputs.cache-hit == 'true' + shell: bash + run: poetry run pip --version >/dev/null 2>&1 || rm -rf .venv + + - name: Check that the lock file is up to date + shell: bash + run: | + poetry lock --check + + - name: Install deps + shell: bash + run: | + poetry install + + - name: Style checking + shell: bash + run: | + poetry run ruff format --check bwapy tests + + - name: Run lint + shell: bash + run: | + poetry run ruff check bwapy tests + + - name: Run mypy + shell: bash + run: | + poetry run mypy bwapy tests --config=pyproject.toml + + - name: Run pytest + shell: bash + run: | + poetry run python -m pytest --cov=bwapy --cov-report=xml --cov-branch + + - name: Run docs + shell: bash + run: | + set -euo pipefail + poetry run mkdocs build --strict + + - name: Upload code coverage + uses: codecov/codecov-action@v4.5.0 + with: + token: ${{ secrets.CODECOV_TOKEN }} diff --git a/.github/workflows/readthedocs.yaml b/.github/workflows/readthedocs.yaml new file mode 100644 index 0000000..7625296 --- /dev/null +++ b/.github/workflows/readthedocs.yaml @@ -0,0 +1,20 @@ +name: readthedocs/actions +on: + pull_request_target: + types: + - opened + # Execute this action only on PRs that touch + # documentation files. + paths: + - "docs/**" + +permissions: + pull-requests: write + +jobs: + documentation-links: + runs-on: ubuntu-24.04 + steps: + - uses: readthedocs/actions/preview@v1 + with: + project-slug: "bwapy" diff --git a/.readthedocs.yml b/.readthedocs.yml new file mode 100644 index 0000000..04913cd --- /dev/null +++ b/.readthedocs.yml @@ -0,0 +1,12 @@ +version: 2 +build: + os: ubuntu-22.04 + tools: + python: "3.11" + jobs: + post_install: + - pip install poetry==1.8.3 + - poetry config virtualenvs.create false + - VIRTUAL_ENV=$READTHEDOCS_VIRTUALENV_PATH poetry install +mkdocs: + configuration: mkdocs.yml diff --git a/README.md b/README.md index 3ef54b4..15567d9 100644 --- a/README.md +++ b/README.md @@ -1 +1,29 @@ +[![Language][language-badge]][language-link] +[![Code Style][code-style-badge]][code-style-link] +[![Type Checked][type-checking-badge]][type-checking-link] +[![PEP8][pep-8-badge]][pep-8-link] +[![Code Coverage][code-coverage-badge]][code-coverage-link] +[![License][license-badge]][license-link] + + +[language-badge]: http://img.shields.io/badge/language-python-brightgreen.svg +[language-link]: http://www.python.org/ +[code-style-badge]: https://img.shields.io/badge/code%20style-black-000000.svg +[code-style-link]: https://black.readthedocs.io/en/stable/ +[type-checking-badge]: http://www.mypy-lang.org/static/mypy_badge.svg +[type-checking-link]: http://mypy-lang.org/ +[pep-8-badge]: https://img.shields.io/badge/code%20style-pep8-brightgreen.svg +[pep-8-link]: https://www.python.org/dev/peps/pep-0008/ +[code-coverage-badge]: https://codecov.io/gh/fulcrumgenomics/bwapy/branch/main/graph/badge.svg +[code-coverage-link]: https://codecov.io/gh/fulcrumgenomics/bwapy +[license-badge]: http://img.shields.io/badge/license-MIT-blue.svg +[license-link]: https://github.com/fulcrumgenomics/bwapy/blob/main/LICENSE + # bwapy + +Python language bindings for [bwa][https://github.com/lh3/bwa]. + +See documentation on [bwapy.readthedocs.org][rtd-link]. + + +[rtd-link]: http://bwapy.readthedocs.org/en/stable diff --git a/build.py b/build.py new file mode 100644 index 0000000..4d80270 --- /dev/null +++ b/build.py @@ -0,0 +1,112 @@ +from setuptools import Extension, Distribution +from typing import List + +from Cython.Build import cythonize +from Cython.Distutils.build_ext import new_build_ext as cython_build_ext +import multiprocessing +from pathlib import Path + +SOURCE_DIR = Path("bwapy") +BUILD_DIR = Path("cython_build") +compile_args = [] +link_args = [] +include_dirs = ["bwa"] +libraries = ['m', 'z', 'pthread'] +library_dirs=['bwa'] +extra_objects = [] #glob.glob(os.path.join('bwa', '*.o')) +h_files = [] +c_files = [] +for root_dir in ["bwa", "bwapy"]: + h_files.extend(str(x) for x in Path(root_dir).rglob("*.h")) + c_files.extend(str(x) for x in Path(root_dir).rglob("*.c") if x.name not in ['example.c', 'main.c']) + +extension_module = Extension( + name='bwapy.libbwapy', + sources=['bwapy/libbwapy.pyx'] + c_files, + depends=h_files, + extra_compile_args=compile_args, + extra_link_args=link_args, + extra_objects=extra_objects, + include_dirs=include_dirs, + language='c', + libraries=libraries, + library_dirs=library_dirs, +) + + +def cythonize_helper(extension_modules: List[Extension]) -> List[Extension]: + """Cythonize all Python extensions""" + + return cythonize( + module_list=extension_modules, + + # Don't build in source tree (this leaves behind .c files) + build_dir=BUILD_DIR, + + # Don't generate an .html output file. Would contain source. + annotate=False, + + # Parallelize our build + nthreads=multiprocessing.cpu_count() * 2, + + # Tell Cython we're using Python 3. Becomes default in Cython 3 + compiler_directives={"language_level": "3"}, + + # (Optional) Always rebuild, even if files untouched + force=True, + ) + +CLASSIFIERS = ''' +Development Status :: 4 - Beta +Intended Audience :: Science/Research +Intended Audience :: Developers +License :: OSI Approved +Programming Language :: Python +Topic :: Software Development +Topic :: Scientific/Engineering +Operating System :: POSIX +Operating System :: Unix +Operating System :: MacOS +''' + + +def build(): + # Collect and cythonize all files + extension_modules = cythonize_helper([extension_module]) + + # Use Setuptools to collect files + distribution = Distribution({ + "name": "bwapy", + 'version': '0.0.1', # FIXME + 'description': 'Todo', # FIXME + 'long_description': 'FIXME', + 'long_description_content_type': 'text/x-rst', + 'author': 'Nils Homer', + 'author_email': 'nils@fulcrumgenomics.com', + 'license': 'MIT', + 'platforms': ['POSIX', 'UNIX', 'MacOS'], + 'classifiers': [_f for _f in CLASSIFIERS.split('\n') if _f], + 'url': 'https://github.com/fulcrumgenomics/bwapy', + 'packages': ['bwapy'], + 'package_dirs': {'bwapy': 'bwapy'}, + "ext_modules": extension_modules, + "cmdclass": { + "build_ext": cython_build_ext, + }, + }) + + # Grab the build_ext command and copy all files back to source dir. + # Done so Poetry grabs the files during the next step in its build. + build_ext_cmd = distribution.get_command_obj("build_ext") + build_ext_cmd.ensure_finalized() + # Set the value to 1 for "inplace", with the goal to build extensions + # in build directory, and then copy all files back to the source dir + # (under the hood, "copy_extensions_to_source" will be called after + # building the extensions). This is done so Poetry grabs the files + # during the next step in its build. + build_ext_cmd.inplace = 1 + build_ext_cmd.run() + + +if __name__ == "__main__": + build() diff --git a/bwapy.yml b/bwapy.yml new file mode 100644 index 0000000..768f735 --- /dev/null +++ b/bwapy.yml @@ -0,0 +1,9 @@ +name: bwapy +channels: + - defaults + - conda-forge + - bioconda +dependencies: + - python=3.11 + - cython=3.0.11 + - pysam=0.22.1 diff --git a/bwapy/__init__.py b/bwapy/__init__.py new file mode 100644 index 0000000..6fc987c --- /dev/null +++ b/bwapy/__init__.py @@ -0,0 +1,4 @@ +import bwapy.libbwapy as libbwapy +from bwapy.libbwapy import * # noqa: F403 + +__all__ = libbwapy diff --git a/bwapy/libbwapy.pxd b/bwapy/libbwapy.pxd new file mode 100644 index 0000000..ee5e6dd --- /dev/null +++ b/bwapy/libbwapy.pxd @@ -0,0 +1,119 @@ +# cython: language_level=3 + +from libc.stdint cimport uint8_t, uint64_t, uint16_t, uint32_t, int64_t, int32_t +from libc.stdio cimport FILE + +cdef extern from "libbwapy_utils.h": + void bwa_cal_pac_pos_with_bwt(const bntseq_t *bns, int n_seqs, bwa_seq_t *seqs, int max_mm, + float fnr, bwt_t *bwt) + +cdef extern from "utils.h": + int err_fseek(FILE *stream, long offset, int whence) + size_t err_fread_noeof(void *ptr, size_t size, size_t nmemb, FILE *stream) + +cdef extern from "bntseq.h": + unsigned char nst_nt4_table[256] + int bns_cnt_ambi(const bntseq_t *bns, int64_t pos_f, int len, int *ref_id) + +cdef extern from "bwa.h": + char * bwa_idx_infer_prefix(const char * hint) + +cdef extern from "bwt.h": + ctypedef struct bwt_t: + int sa_intv + + bwt_t *bwt_restore_bwt(const char *fn) + void bwt_restore_sa(const char *fn, bwt_t *bwt); + void bwt_destroy(bwt_t *bwt) + +cdef extern from "bwtaln.h": + int BWA_TYPE_NO_MATCH + int BWA_MODE_LOGGAP + int BWA_MODE_GAPE + + int __cigar_op(uint16_t __cigar) + int __cigar_len(uint16_t __cigar) + + ctypedef struct gap_opt_t: + int trim_qual + int s_mm + int s_gapo + int s_gape + int mode # bit 24-31 are the barcode length + int indel_end_skip + int max_del_occ + int max_entries + float fnr + int max_diff + int max_gapo + int max_gape + int max_seed_diff + int seed_len + int n_threads + int max_top2 + int trim_qual + int sam + char *rg_line + int n_occ + int interactive_mode + int with_md + + gap_opt_t *gap_init_opt() + void gap_print_opt(const gap_opt_t *opt) + + void seq_reverse(int len, unsigned char *seq, int is_comp) + + ctypedef struct bwt_aln1_t: + pass + +cdef extern from "bntseq.h": + ctypedef struct bntann1_t: + int64_t offset + int32_t len + char *name + + ctypedef struct bntseq_t: + int64_t l_pac + bntann1_t *anns + FILE * fp_pac + + bntseq_t * bns_restore(const char * prefix) + void bns_destroy(bntseq_t *bns) + +cdef extern from "kseq.h": + ctypedef struct kstring_t: + char *s + +cdef extern from "bwase.h": + void bwa_aln2seq_core(int n_aln, const bwt_aln1_t *aln, bwa_seq_t *s, int set_main, int n_multi) + int64_t pos_end(const bwa_seq_t *p) + void bwa_refine_gapped(const bntseq_t *bns, int n_seqs, bwa_seq_t *seqs, unsigned char *_pacseq) + char *bwa_cal_md1(int n_cigar, uint16_t *cigar, int len, uint64_t pos, unsigned char *seq, uint64_t l_pac, unsigned char *pacseq, kstring_t *str, int *_nm) + void bwase_initialize() + +cdef extern from "bwtaln.h": + ctypedef struct bwa_seq_t: + + char *name + uint8_t *seq + uint8_t *rseq + uint8_t *qual + uint32_t len + uint32_t strand + uint32_t type + int mapQ + int clip_len + bwt_aln1_t *aln + int n_aln + uint16_t pos + uint16_t *cigar + int n_cigar + int tid + uint32_t full_len + uint32_t nm + char *md + + + void bwa_free_read_seq(int n_seqs, bwa_seq_t *seqs) + + void bwa_cal_sa_reg_gap(int tid, bwt_t *const bwt, int n_seqs, bwa_seq_t *seqs, const gap_opt_t *opt) diff --git a/bwapy/libbwapy.pyi b/bwapy/libbwapy.pyi new file mode 100644 index 0000000..977e360 --- /dev/null +++ b/bwapy/libbwapy.pyi @@ -0,0 +1,42 @@ +from pathlib import Path +from typing import List + +from pysam import AlignedSegment +from pysam import AlignmentHeader +from pysam import FastxRecord + +class BwaAlnOptions: + max_hits: int + def __init__(self, max_hits: int = 3) -> None: ... + +class BwaAlnOptionsBuilder: + _options: BwaAlnOptions + def __init__(self, options: BwaAlnOptions | None = None) -> None: ... + def build(self) -> BwaAlnOptions: ... + def max_mismatches(self, value: int) -> BwaAlnOptionsBuilder: ... # -n + # def fnr(self, value: float) -> BwaOptionsBuilder: ... # -n + def max_gap_opens(self, value: int) -> BwaAlnOptionsBuilder: ... # -o + def max_gap_extensions(self, value: int) -> BwaAlnOptionsBuilder: ... # -e + def min_indel_to_end_distance(self, value: int) -> BwaAlnOptionsBuilder: ... # -i + def max_occurences_for_extending_long_deletion( + self, value: int + ) -> BwaAlnOptionsBuilder: ... # -d + def seed_length(self, value: int) -> BwaAlnOptionsBuilder: ... # -l + def max_mismatches_in_seed(self, value: int) -> BwaAlnOptionsBuilder: ... # -k + def mismatch_penalty(self, value: int) -> BwaAlnOptionsBuilder: ... # -M + def gap_open_penalty(self, value: int) -> BwaAlnOptionsBuilder: ... # -O + def gap_extension_penalty(self, value: int) -> BwaAlnOptionsBuilder: ... # -E + def stop_at_max_best_hits(self, value: int) -> BwaAlnOptionsBuilder: ... # -R + def max_hits(self, value: int) -> BwaAlnOptionsBuilder: ... # bwa samse -n + def log_scaled_gap_penalty(self, value: bool = True) -> BwaAlnOptionsBuilder: ... # -L + +ERROR_HANDLER: str +TEXT_ENCODING: str + +class BwaIndex: + header: AlignmentHeader + def __init__(self, prefix: str | Path) -> None: ... + +class BwaAln: + def __init__(self, prefix: str | Path | None = None, index: BwaIndex | None = None) -> None: ... + def align(self, opt: BwaAlnOptions, queries: List[FastxRecord]) -> List[AlignedSegment]: ... diff --git a/bwapy/libbwapy.pyx b/bwapy/libbwapy.pyx new file mode 100644 index 0000000..97c4862 --- /dev/null +++ b/bwapy/libbwapy.pyx @@ -0,0 +1,356 @@ +# cython: language_level=3 + +from pathlib import Path +from typing import List + +from cpython cimport PyBytes_Check, PyUnicode_Check +from libc.stdint cimport uint8_t +from libc.stdio cimport SEEK_SET +from libc.stdlib cimport calloc, free +from libc.string cimport strncpy, strcat +from pysam import FastxRecord, AlignmentFile, AlignedSegment + +__all__ = [ + "BwaAlnOptions", + "BwaAlnOptionsBuilder", + "BwaIndex", + "BwaAln", +] + +cdef class BwaAlnOptions: + """The container for options for [`BwaAln`][bwapy.BwaAln]. + + Use [`BwaAlnOptionsBuilder`][bwapy.BwaAlnOptionsBuilder] to use and set custom options. + """ + cdef gap_opt_t * _delegate + _max_hits: int + + def __init__(self, max_hits: int = 3): + self._max_hits = max_hits + self._cinit() + + cdef _cinit(self): + self._delegate = gap_init_opt() + + def __dealloc__(self): + free(self._delegate) + + +cdef class BwaAlnOptionsBuilder: + """Builder for options for [`BwaAln`][bwapy.BwaAln]""" + _options: BwaAlnOptions + + def __init__(self, options: BwaAlnOptions | None = None) -> None: + self._options: BwaAlnOptions = BwaAlnOptions() if options is None else options + + def build(self) -> BwaAlnOptions: + return self._options + + cdef max_mismatches(self, value: int): + """bwa aln -n """ + self._options._delegate.s_mm = value + return self + #cdef fnr(self, value: float) -> BwaOptionsBuilder: ... # -n + cdef max_gap_opens(self, value: int): + """bwa aln -o """ + self._options._delegate.max_gapo = value + return self + + cdef max_gap_extensions(self, value: int): + """bwa aln -e """ + self._options._delegate.max_gape = value + if self._delegate.max_gape > 0: + self._delegate.mode &= ~BWA_MODE_GAPE + return self + + cdef min_indel_to_end_distance(self, value: int): + """bwa aln -i """ + self._options._delegate.indel_end_skip = value + return self + + def max_occurences_for_extending_long_deletion(self, value: int): + """bwa aln -d """ + self._options._delegate.max_del_occ = value + return self + + def seed_length(self, value: int): + """bwa aln -l """ + self._options._delegate.seed_len = value + return self + + def max_mismatches_in_seed(self, value: int): + """bwa aln -k """ + self._options._delegate.max_seed_diff = value + return self + + def mismatch_penalty(self, value: int): + """bwa aln -M """ + self._options._delegate.s_mm = value + return self + + def gap_open_penalty(self, value: int): + """bwa aln -O """ + self._options._delegate.s_gapo = value + return self + + def gap_extension_penalty(self, value: int): + """bwa aln -E """ + self._options._delegate.s_gape = value + return self + + def stop_at_max_best_hits(self, value: int): + """bwa aln -R """ + self._options._delegate.max_top2 = value + return self + + def max_hits(self, value: int): + """bwa samse -n """ + self._options._max_hits = value + return self + + def log_scaled_gap_penalty(self, value: bool = True): + """bwa aln -L""" + if value: + self._options._delegate.mode |= BWA_MODE_LOGGAP + else: + self._options._delegate.mode &= ~BWA_MODE_LOGGAP + return self + + +cdef str ERROR_HANDLER = 'strict' +cdef str TEXT_ENCODING = 'utf-8' + + +cdef bytes force_bytes(object s, encoding=None, errors=None): + """convert string or unicode object to bytes, assuming + utf8 encoding. + """ + if s is None: + return None + elif PyBytes_Check(s): + return s + elif PyUnicode_Check(s): + return s.encode(encoding or TEXT_ENCODING, errors or ERROR_HANDLER) + else: + raise TypeError("Argument must be string, bytes or unicode.") + + + +cdef class BwaIndex: + """Contains the index and nucleotide sequence for Bwa""" + cdef bwt_t *_bwt + cdef bntseq_t *_bns + cdef public object header + + def __init__(self, prefix: str | Path): + self._cinit(f"{prefix}") + + cdef _cinit(self, prefix): + cdef char *local_prefix + + prefix = bwa_idx_infer_prefix(force_bytes(prefix)) + if not prefix: + # FIXME: better error message + raise Exception("Could not find the index") + + # the SAM header from the sequence dictionary + seq_dict = Path(prefix.decode("utf-8")).with_suffix(".dict") + # TODO: error message when seq_dict is missing? + with seq_dict.open("r") as fh: + with AlignmentFile(fh) as reader: + self.header = reader.header + + # from bwaaln.c + local_prefix = calloc(sizeof(char), len(prefix) + 10) + + # load BWT + strncpy(local_prefix, prefix, len(prefix)) + strcat(local_prefix, force_bytes(".bwt")) + self._bwt = bwt_restore_bwt(local_prefix) + + # load SA + local_prefix[len(prefix)] = b'\0' # trim off .bwt first + strcat(local_prefix, force_bytes(".sa")) + bwt_restore_sa(local_prefix, self._bwt) + + # load BNS + self._bns = bns_restore(prefix) + + def __dealloc__(self): + bwt_destroy(self._bwt) + self._bwt = NULL + bns_destroy(self._bns) + self._bns = NULL + + +cdef class BwaAln: + """The class to align reads with `bwa aln`.""" + + cdef BwaIndex _index + cdef unsigned char* _pacseq + + def __init__(self, prefix: str | Path | None = None, index: BwaIndex | None = None): + if prefix is not None: + assert Path(prefix).exists() + self._index = BwaIndex(prefix=prefix) + elif index is not None: + self._index = index + else: + raise Exception("Either prefix or index must be given") + + bwase_initialize() + + # initialize the packed binary reference sequence + self._pacseq = calloc(self._index._bns.l_pac//4+1, 1) + err_fseek(self._index._bns.fp_pac, 0, SEEK_SET) + err_fread_noeof(self._pacseq, 1, self._index._bns.l_pac//4+1, self._index._bns.fp_pac) + + # TODO: a list of records... + def align(self, opt: BwaAlnOptions, queries: List[FastxRecord]) -> List[AlignedSegment]: + """Align one or more queries with `bwa aln`. + + Args: + opt: the alignment options + queries: the queries to align + + Returns: + one alignment per query + """ + return self._calign(opt, queries) + + cdef _copy_seq(self, q: FastxRecord, bwa_seq_t* s): + seq_len = len(q.sequence) + s.len = seq_len + s.clip_len = seq_len + s.full_len = seq_len + s.seq = calloc(sizeof(uint8_t), seq_len + 1) + s.rseq = calloc(sizeof(uint8_t), seq_len + 1) + s.qual = calloc(sizeof(uint8_t), seq_len + 1) + + # use seq_reverse from bwaseqio.c + for i, base in enumerate(q.sequence): + s.seq[i] = nst_nt4_table[ord(base)] + s.qual[i] = 40 # FIXME: parameterize + s.seq[seq_len] = b'\0' + s.qual[seq_len] = b'\0' + seq_reverse(seq_len, s.seq, + 0) # // *IMPORTANT*: will be reversed back in bwa_refine_gapped() + seq_reverse(seq_len, s.rseq, 0) + + s.name = calloc(sizeof(char), len(q.name) + 1) + strncpy(s.name, force_bytes(q.name), len(q.name)) + s.name[len(q.name)] = b'\0' + + cdef _build_alignment(self, query: FastxRecord, bwa_seq_t *seq, kstring_t *kstr): + cdef int reference_id + cdef int nm + + # make a default, unmapped, empty record + rec = AlignedSegment(header=self._index.header) + rec.query_name = query.name + rec.reference_id = -1 + rec.reference_start = -1 + rec.mapping_quality = 0 + rec.is_paired = False + rec.is_read1 = True + rec.is_read2 = False + rec.is_qcfail = False + rec.is_duplicate = False + rec.is_secondary = False + rec.is_supplementary = False + rec.is_unmapped = True + rec.query_sequence = query.sequence + rec.query_qualities = query.quality + + if seq.type == BWA_TYPE_NO_MATCH: # unmapped read + # TODO: custom bwa tags: RG, BC, XC + return rec + + ref_len_in_alignment = pos_end(seq) - seq.pos + + # if on the reverse strand, reverse the query sequence and qualities + rec.query_sequence = query.sequence[::-1] + if query.quality is not None: + rec.query_qualities = query.quality[::-1] + + # reference id + nn = bns_cnt_ambi(self._index._bns, seq.pos, ref_len_in_alignment, &reference_id) + rec.reference_id = reference_id + + # make this unmapped if we map off the end of the contig + rec.is_unmapped = seq.pos + ref_len_in_alignment - self._index._bns.anns[ + rec.reference_id].offset > self._index._bns.anns[rec.reference_id].len + + # strand, reference start, and mapping quality + if seq.strand: + rec.is_reverse = True + rec.reference_start = seq.pos - self._index._bns.anns[rec.reference_id].offset + 1 + rec.mapping_quality = seq.mapQ + + # cigar + cigar = "" + if seq.cigar: + for j in range(seq.n_cigar): + cigar_len = __cigar_len(seq.cigar[j]) + cigar_op = "MIDS"[__cigar_op(seq.cigar[j])] + cigar = f"{cigar}{cigar_len}{cigar_op}" + elif seq.type != BWA_TYPE_NO_MATCH: + cigar = f"{seq.len}M" + rec.cigarstring = cigar + + # # tags + if seq.type != BWA_TYPE_NO_MATCH: + attrs = dict() + attrs["MD"] = f"{seq.md}" + attrs["NM"] = f"{seq.nm}" + rec.set_tags(list(attrs.items())) + # # TODO:the custom bwa tags: XT, NM, XN, SM, AM, X0, X1, XM, XO, XG, XA, HN + + return rec + + cdef _calign(self, opt: BwaAlnOptions, queries: List[FastxRecord]): + cdef bwa_seq_t* seqs + cdef bwa_seq_t* s + cdef char* s_char + cdef kstring_t* kstr + + kstr = calloc(sizeof(kstring_t), 1) + + # copy FastqProxy into bwa_seq_t + num_seqs = len(queries) + seqs = calloc(sizeof(bwa_seq_t), num_seqs) + for i in range(num_seqs): + self._copy_seq(queries[i], &seqs[i]) + seqs[i].tid = -1 + + # this is `bwa aln`, and the rest is `bwa samse` + bwa_cal_sa_reg_gap(0, self._index._bwt, num_seqs, seqs, opt._delegate) + + # create the full alignment + for i in range(num_seqs): + s = &seqs[i] + # bwa_cal_sa_reg_gap frees name, seq, rseq, and qual, so add them back in again + self._copy_seq(queries[i], s) + bwa_aln2seq_core(s.n_aln, s.aln, s, 1, opt._max_hits) + + # calculate the genomic position given the suffix array offsite + bwa_cal_pac_pos_with_bwt(self._index._bns, num_seqs, seqs, opt._delegate.max_diff, opt._delegate.fnr, self._index._bwt) + + # TODO: can `_pacseq` be passed here? + # refine gapped alignment + bwa_refine_gapped(self._index._bns, num_seqs, seqs, NULL) + + # create the AlignedSegment from FastxRecrod and bwa_seq_t. + recs = [ + self._build_alignment(query=queries[i], seq=&seqs[i], kstr=kstr) + for i in range(num_seqs) + ] + + bwa_free_read_seq(num_seqs, seqs) + free(kstr.s) + free(kstr) + + return recs + + def __dealloc__(self) -> None: + free(self._pacseq) \ No newline at end of file diff --git a/bwapy/libbwapy_utils.c b/bwapy/libbwapy_utils.c new file mode 100644 index 0000000..b3c2d27 --- /dev/null +++ b/bwapy/libbwapy_utils.c @@ -0,0 +1,25 @@ +#include "bntseq.h" +#include "bwt.h" +#include "bwtaln.h" +#include "kstring.h" +#include "libbwapy_utils.h" +#include "bwase.h" + + +void bwa_cal_pac_pos_with_bwt(const bntseq_t *bns, int n_seqs, bwa_seq_t *seqs, int max_mm, float fnr, bwt_t *bwt) +{ + int i, j, strand, n_multi; + char str[1024]; + for (i = 0; i != n_seqs; ++i) { + bwa_seq_t *p = &seqs[i]; + bwa_cal_pac_pos_core(bns, bwt, p, max_mm, fnr); + for (j = n_multi = 0; j < p->n_multi; ++j) { + bwt_multi1_t *q = p->multi + j; + q->pos = bwa_sa2pos(bns, bwt, q->pos, p->len + q->ref_shift, &strand); + q->strand = strand; + if (q->pos != p->pos && q->pos != (bwtint_t)-1) + p->multi[n_multi++] = *q; + } + p->n_multi = n_multi; + } +} \ No newline at end of file diff --git a/bwapy/libbwapy_utils.h b/bwapy/libbwapy_utils.h new file mode 100644 index 0000000..cfd6769 --- /dev/null +++ b/bwapy/libbwapy_utils.h @@ -0,0 +1,8 @@ +#include "bntseq.h" +#include "bwt.h" +#include "bwtaln.h" +#include "kstring.h" + + +// Version of "bwa_cal_pac_pos" bwase.c that can use an already loaded forward suffix array (BWT) +void bwa_cal_pac_pos_with_bwt(const bntseq_t *bns, int n_seqs, bwa_seq_t *seqs, int max_mm, float fnr, bwt_t *bwt); diff --git a/bwapy/py.typed b/bwapy/py.typed new file mode 100644 index 0000000..e69de29 diff --git a/codecov.yml b/codecov.yml new file mode 100644 index 0000000..a901cc2 --- /dev/null +++ b/codecov.yml @@ -0,0 +1,2 @@ +ignore: + - "**/test_*.py" # don't compute coverage of tests diff --git a/docs/index.md b/docs/index.md new file mode 100644 index 0000000..01ca5f5 --- /dev/null +++ b/docs/index.md @@ -0,0 +1,9 @@ +# bwapy + +Python bindings for BWA + +## Documentation Contents + +* [Installation](installation-and-developers-documentation.md) +* [API](reference/bwapy/index.md) + diff --git a/docs/installation-and-developers-documentation.md b/docs/installation-and-developers-documentation.md new file mode 100644 index 0000000..b69465a --- /dev/null +++ b/docs/installation-and-developers-documentation.md @@ -0,0 +1,76 @@ +# Installation and Developer's Documentation + +## Recommended Installation + +The package `bwapy` requires cloning [`bwa`](https://github.com/lh3/bwa) into the source directory. + +```console +git clone git@github.com:lh3/bwa.git +``` + +## Installation for Development and Release + +Install the Python build tool [`poetry`](https://python-poetry.org/docs/#installing-with-the-official-installer) + +Configure `poetry` to install into pre-existing virtual environments: + +```console +poetry config virtualenvs.create false +``` + +```console +poetry install +``` + +## Checking the Build + +Use `poetry` to test your code. + +```console +poetry run pytest +``` + +Note that `poetry run pytest` will run `mypy` checks, `ruff` checks, `pytest` unit tests, and will provide a unit test coverage report. +However, `pytest` will neither run the ruff formatter nor apply `ruff`'s automatic lint fixes, which can be done by calling `ruff` directly. + +```console +poetry run ruff format && poetry run ruff check --fix +``` + +## Building the Documentation + +Use `mkdocs` to build and serve the documentation. + +```console +poetry run mkdocs build && poetry run mkdocs serve +``` + +## Creating a Release on PyPi + +1. Clone the repository recursively and ensure you are on the `main` (un-dirty) branch +2. Checkout a new branch to prepare the library for release +3. Bump the version of the library to the desired SemVer with `poetry version #.#.#` +4. Commit the version bump changes with a Git commit message like `chore(release): bump to #.#.#` +5. Push the commit to the upstream remote, open a PR, ensure tests pass, and seek reviews +6. Squash merge the PR +7. Tag the new commit on the main branch of the origin repository with the new SemVer + +> [!NOTE] +> This project follows [Semantic Versioning](https://semver.org/). +> In brief: +> +> * `MAJOR` version when you make incompatible API changes +> * `MINOR` version when you add functionality in a backwards compatible manner +> * `PATCH` version when you make backwards compatible bug fixes + +GitHub Actions will take care of the remainder of the deployment and release process with: + +1. Unit tests will be run for safety-sake +2. A source distribution will be built +3. Multi-arch multi-Python binary distributions will be built +4. Assets will be deployed to PyPi with the new SemVer +5. A [Conventional Commit](https://www.conventionalcommits.org/en/v1.0.0/)-aware changelog will be drafted +6. A GitHub release will be created with the new SemVer and the drafted changelog + +> [!IMPORTANT] +> Consider editing the changelog if there are any errors or necessary enhancements. diff --git a/docs/scripts/gen_ref_pages.py b/docs/scripts/gen_ref_pages.py new file mode 100644 index 0000000..97b1411 --- /dev/null +++ b/docs/scripts/gen_ref_pages.py @@ -0,0 +1,49 @@ +"""Generate the code reference pages and navigation.""" + +# Adapted from https://mkdocstrings.github.io/recipes/#bind-pages-to-sections-themselves + +from pathlib import Path + +import mkdocs_gen_files + +nav = mkdocs_gen_files.Nav() + +library_name = "bwapy" +root = Path(__file__).parent.parent.parent +src = root +module_root = root / library_name + +assert module_root.exists(), ( + f"module root does not exist: {module_root}\n" + f"Are you sure the library name '{library_name}' is correct?" +) + +for path in sorted(module_root.rglob("*.py")): + module_path = path.relative_to(src).with_suffix("") + doc_path = path.relative_to(src).with_suffix(".md") + full_doc_path = Path("reference", doc_path) + + parts = tuple(module_path.parts) + + # ignore tests + if "tests" in parts: + continue + + if parts[-1] == "__init__": + parts = parts[:-1] + doc_path = doc_path.with_name("index.md") + full_doc_path = full_doc_path.with_name("index.md") + elif parts[-1] == "__main__": + continue + + nav[parts] = doc_path.as_posix() + + with mkdocs_gen_files.open(full_doc_path, "w") as fd: + ident = ".".join(parts) + fd.write(f"::: {ident}") + + #mkdocs_gen_files.set_edit_path(full_doc_path, Path("../") / path.relative_to(root)) + mkdocs_gen_files.set_edit_path(full_doc_path, path.relative_to(root)) + +with mkdocs_gen_files.open("reference/SUMMARY.md", "w") as nav_file: + nav_file.writelines(nav.build_literate_nav()) diff --git a/mkdocs.yml b/mkdocs.yml new file mode 100644 index 0000000..33a05b0 --- /dev/null +++ b/mkdocs.yml @@ -0,0 +1,52 @@ +site_name: bwapy +site_url: https://bwapy.readthedocs.io/en/latest/ +use_directory_urls: false +theme: + name: material + highlightjs: true + hljs_languages: + - python + palette: + primary: teal + navstyle: dark + include_sidebar: true + collapse_navigation: false +repo_url: https://github.com/fulcrumgenomics/bwapy +plugins: + - autorefs: + resolve_closest: true + - include-markdown + - search + - gen-files: + scripts: + - docs/scripts/gen_ref_pages.py + - literate-nav: + nav_file: SUMMARY.md + - section-index + - mkdocstrings: + handlers: + python: + options: + docstring_section_style: table + docstring_style: google + group_by_category: true + separate_signature: true + show_category_heading: true + show_if_no_docstring: false + show_root_toc_entry: true + show_signature_annotations: true + signature_crossrefs: true + show_submodules: true + - table-reader +markdown_extensions: + - pymdownx.highlight: + anchor_linenums: true + line_spans: __span + pygments_lang_class: true + - pymdownx.inlinehilite + - pymdownx.snippets + - pymdownx.superfences + - toc: + permalink: true +exclude_docs: | + test_*.py diff --git a/poetry.lock b/poetry.lock new file mode 100644 index 0000000..2f27909 --- /dev/null +++ b/poetry.lock @@ -0,0 +1,1648 @@ +# This file is automatically @generated by Poetry 1.4.0 and should not be changed by hand. + +[[package]] +name = "babel" +version = "2.16.0" +description = "Internationalization utilities" +category = "dev" +optional = false +python-versions = ">=3.8" +files = [ + {file = "babel-2.16.0-py3-none-any.whl", hash = "sha256:368b5b98b37c06b7daf6696391c3240c938b37767d4584413e8438c5c435fa8b"}, + {file = "babel-2.16.0.tar.gz", hash = "sha256:d1f3554ca26605fe173f3de0c65f750f5a42f924499bf134de6423582298e316"}, +] + +[package.extras] +dev = ["freezegun (>=1.0,<2.0)", "pytest (>=6.0)", "pytest-cov"] + +[[package]] +name = "bracex" +version = "2.5.post1" +description = "Bash style brace expander." +category = "dev" +optional = false +python-versions = ">=3.8" +files = [ + {file = "bracex-2.5.post1-py3-none-any.whl", hash = "sha256:13e5732fec27828d6af308628285ad358047cec36801598368cb28bc631dbaf6"}, + {file = "bracex-2.5.post1.tar.gz", hash = "sha256:12c50952415bfa773d2d9ccb8e79651b8cdb1f31a42f6091b804f6ba2b4a66b6"}, +] + +[[package]] +name = "certifi" +version = "2024.8.30" +description = "Python package for providing Mozilla's CA Bundle." +category = "dev" +optional = false +python-versions = ">=3.6" +files = [ + {file = "certifi-2024.8.30-py3-none-any.whl", hash = "sha256:922820b53db7a7257ffbda3f597266d435245903d80737e34f8a45ff3e3230d8"}, + {file = "certifi-2024.8.30.tar.gz", hash = "sha256:bec941d2aa8195e248a60b31ff9f0558284cf01a52591ceda73ea9afffd69fd9"}, +] + +[[package]] +name = "charset-normalizer" +version = "3.4.0" +description = "The Real First Universal Charset Detector. 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+from bwapy import BwaIndex + + +@pytest.fixture() +def ref_fasta() -> Path: + cur_dir = Path(__file__).parent + fasta: Path = cur_dir / "data" / "e_coli_k12.fasta" + return fasta + + +def test_bwapy_options() -> None: + BwaAlnOptions() + + +def test_bwapy_options_builder() -> None: + builder = BwaAlnOptionsBuilder() + builder.build() + # TODO: test setting individual options... + + +def test_bwapy_index(ref_fasta: Path) -> None: + BwaIndex(prefix=ref_fasta) + + +def test_bwapy(ref_fasta: Path) -> None: + opt = BwaAlnOptions() + bwa = BwaAln(prefix=ref_fasta) + sequence = "gttacctgccgtgagtaaattaaaattttattgacttaggtcactaaatactttaaccaatataggcatagcgcacagac" + fastqs = [FastxRecord(name="test", sequence=sequence)] + + recs = bwa.align(opt=opt, queries=fastqs) + assert len(recs) == 1 + rec = recs[0] + assert rec.query_name == "test" + assert rec.reference_start == 81 + assert rec.cigarstring == "80M"

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