doc: Expand the documentation for OffTargetDetector

prymer/offtarget/offtarget_detector.py

@@ -102,8 +102,10 @@ class OffTargetResult:
     """Information obtained by running a single primer pair through the off-target detector.
 
     Attributes:
-        primer_pair: the primer submitted
-        passes: True if the primer pair passes all checks, False otherwise
+        primer_pair: the primer pair submitted
+        passes: True if neither primer exceeds the specified maximum number of hits, and the primer
+            pair would generate an acceptable number of amplicons in the reference genome (i.e.
+            `min_primer_pair_hits <= num_amplicons <= max_primer_pair_hits`). False otherwise.
         cached: True if this result is part of a cache, False otherwise.  This is useful for testing
         spans: the set of mappings of the primer pair to the genome or an empty list if mappings
             were not retained
@@ -122,15 +124,28 @@ class OffTargetResult:
 
 
 class OffTargetDetector:
-    """A class for detecting off-target mappings of primers and primer pairs that uses a custom
-    version of "bwa aln".
+    """
+    Detect off-target mappings of primers and primer pairs.
 
-    The off-target detection is faster and more sensitive than traditional isPCR and in addition can
-    correctly detect primers that are repetitive and contain many thousands or millions of mappings
-    to the genome.
+    `OffTargetDetector` uses a [custom, interactive
+    version](https://github.com/fulcrumgenomics/bwa-aln-interactive/) of `bwa aln` to perform
+    off-target detection. This approach is faster and more sensitive than traditional isPCR and in
+    addition can correctly detect primers that are repetitive and contain many thousands or millions
+    of mappings to the genome.
 
     Note that while this class invokes BWA with multiple threads, it is not itself thread-safe.
     Only one thread at a time should invoke methods on this class without external synchronization.
+
+    Off-target detection may be performed for individual primers or primer pairs.
+
+    To detect off-target hits for individual primers, use the `OffTargetDetector.filters()` method,
+    which will remove any primers that have more than the specified number of maximum hits against
+    the reference.
+
+    To detect off-target amplification of primer pairs, use the `OffTargetDetector.check_one()` or
+    `OffTargetDetector.check_all()` methods. These methods screen the individual primers in each
+    pair for off-target hits, and verify that the possible amplicons inferred by the primers'
+    alignments does not exceed the specified maximum number of primer pair hits.
     """
 
     def __init__(
@@ -150,6 +165,19 @@ def __init__(
         executable: str | Path = "bwa",
     ) -> None:
         """
+        Initialize an [[OffTargetDetector]].
+
+        This class accepts a large number of parameters to control the behavior of the off-target
+        detection. The parameters are used in the various aspects of off-target detection as
+        follows:
+
+        1. Alignment (off-target hit detection): `ref`, `executable`, `threads`,
+           `three_prime_region_length`, `max_mismatches_in_three_prime_region`, `max_mismatches`,
+           and `max_primer_hits`.
+        2. Filtering of individual primers: `max_primer_hits`.
+        3. Checking of primer pairs: `max_primer_hits`, `min_primer_pair_hits`,
+           `max_primer_pair_hits`, and `max_amplicon_size`.
+
         Args:
             ref: the reference genome fasta file (must be indexed with BWA)
             max_primer_hits: the maximum number of hits an individual primer can have in the genome
@@ -173,10 +201,10 @@ def __init__(
             max_amplicon_size: the maximum amplicon size to consider amplifiable
             cache_results: if True, cache results for faster re-querying
             threads: the number of threads to use when invoking bwa
-            keep_spans: if True, [[OffTargetResult]] objects will have amplicon spans
+            keep_spans: if True, [[OffTargetResult]] objects will be reported with amplicon spans
                 populated, otherwise not
-            keep_primer_spans: if True, [[OffTargetResult]] objects will have left and right
-                primer spans
+            keep_primer_spans: if True, [[OffTargetResult]] objects will be reported with left and
+                right primer spans
             executable: string or Path representation of the `bwa` executable path
         """
         self._primer_cache: dict[str, BwaResult] = {}
@@ -200,25 +228,61 @@ def __init__(
         self._keep_primer_spans: bool = keep_primer_spans
 
     def filter(self, primers: list[PrimerType]) -> list[PrimerType]:
-        """Filters an iterable of Primers to return only those that have less than
-        `max_primer_hits` mappings to the genome."""
+        """
+        Remove primers that have more than `max_primer_hits` mappings to the genome.
+
+        This method maps each primer to the specified reference with `bwa aln` to search for
+        off-target hits. Note that when the reference includes the sequence from which the primers
+        were design, the on-target hit will be included in the hit count. `max_primer_hits` should
+        be set to at least 1 in this case.
+
+        Args:
+            primers: A list of primers to filter.
+
+        Returns:
+            The input primers, filtered to remove any primers with more than `max_primer_hits`
+            mappings to the genome.
+        """
         results: dict[str, BwaResult] = self.mappings_of(primers)
         return [
             primer for primer in primers if results[primer.bases].hit_count <= self._max_primer_hits
         ]
 
     def check_one(self, primer_pair: PrimerPair) -> OffTargetResult:
-        """Checks a PrimerPair for off-target sites in the genome at which it might amplify."""
+        """
+        Check an individual primer pair for off-target amplification.
+
+        See `OffTargetDetector.check_all()` for details.
+
+        Args:
+            primer_pair: The primer pair to check.
+
+        Returns:
+            The results of the off-target detection.
+            See `OffTargetResult` for details regarding the attributes included in the result.
+        """
         result: dict[PrimerPair, OffTargetResult] = self.check_all([primer_pair])
         return result[primer_pair]
 
     def check_all(self, primer_pairs: list[PrimerPair]) -> dict[PrimerPair, OffTargetResult]:
-        """Checks a collection of primer pairs for off-target sites, returning a dictionary of
-        `PrimerPair`s to `OffTargetResult`.
+        """
+        Check a collection of primer pairs for off-target amplification.
+
+        This method maps each primer to the specified reference with `bwa aln` to search for
+        off-target hits. Possible amplicons are identified from the pairwise combinations of these
+        alignments, up to the specified `max_amplicon_size`.
+
+        Primer pairs are marked as passing if both of the following are true:
+        1. Each primer has no more than `max_primer_hits` alignments to the genome.
+        2. The size of the set of possible amplicons does not exceed `max_primer_pair_hits`, and is
+           at least `min_primer_pair_hits`.
+
+        Args:
+            primer_pairs: The primer pairs to check.
 
         Returns:
-            a Map containing all given primer pairs as keys with the values being the result of
-            off-target checking.
+            A dictionary mapping each checked primer pair to its off-target detection results.
+            See `OffTargetResult` for details regarding the attributes included in each result.
         """
 
         primer_pair_results: dict[PrimerPair, OffTargetResult] = {}
@@ -294,10 +358,27 @@ def _build_off_target_result(
         return result
 
     def mappings_of(self, primers: list[PrimerType]) -> dict[str, BwaResult]:
-        """Function to take a set of primers and map any that are not cached, and return mappings
-        for all of them.  Note: the genomics sequence of the returned mappings are on the opposite
-        strand of that of the strand of the primer.  I.e. we map the complementary bases (reversed)
-        to that of the primer."""
+        """
+        Map primers to the reference genome using `bwa aln`.
+
+        Alignment results may be optionally cached for efficiency. Set `cache_results` to `True`
+        when instantiating the [[OffTargetDetector]] to enable caching.
+
+        Any primers without cached results, or all primers when `cache_results` is `False`, will be
+        mapped to the reference genome using `bwa aln` and the specified alignment parameters.
+
+        **Note**: The reverse complement of each primer sequence is used for mapping. The `query`
+        sequence in each `BwaResult` will match the input primer sequence, as will the sequences
+        used as keys in the output dictionary. However, the coordinates reported in each `BwaHit`
+        associated with a result will correspond to complementary sequence.
+
+        Args:
+            primers: A list of primers to map.
+
+        Returns:
+            A dictionary mapping each primer's sequence to its alignment results.
+            See `BwaResult` for details regarding the attributes included in each result.
+        """
 
         primers_to_map: list[PrimerType]
         if not self._cache_results: