countmut 0.3.1__tar.gz → 0.3.2__tar.gz

{countmut-0.3.1/countmut.egg-info → countmut-0.3.2}/PKG-INFO

@@ -1,6 +1,6 @@
 Metadata-Version: 2.4
 Name: countmut
-Version: 0.3.1
+Version: 0.3.2
 Summary: Unified ultra-fast strand-aware mutation counter (C backend + samtools-style -e/-p filters)
 Author-email: Ye Chang <yech1990@gmail.com>
 License-Expression: MIT

{countmut-0.3.1 → countmut-0.3.2}/backend/countmut_core.c

@@ -569,10 +569,11 @@ static void count_interval(worker_t *w, const cm_config *cfg, bam_hdr_t *hdr, FI
             int s = bio_strand(b);
             uint8_t nt = bam_seqi(bam_get_seq(b), p->qpos);
             int base_i = nt16_index(nt);
-            /* Minus reads: stored SEQ is 5'->3' (SAM spec) but qpos walks
-             * CIGAR order (left->right); complement the base into the
-             * reference frame (parity with countmut 0.0.x + pysam pairs). */
-            if (s == 1 && base_i < 4) base_i = 3 - base_i;
+            /* Reverse-oriented reads: stored SEQ is 5'->3' (SAM spec) but qpos
+             * walks CIGAR order (left->right); complement the base into the
+             * reference frame.  Key on bam_is_rev (read orientation), not the
+             * biological strand s (which differs for R2 in paired data). */
+            if (bam_is_rev(b) && base_i < 4) base_i = 3 - base_i;
             /* router-assigned category slot (0..CM_CAT_MAX-1); g_a set in
              * the selection loop for every kept candidate. */
             int cat = w->g_a[i];
@@ -764,10 +765,13 @@ static rw_w *rw_add_base(worker_t *w, const cm_config *cfg, bam_hdr_t *hdr, int
     }
     uint8_t nt = bam_seqi(bam_get_seq(b), qpos);
     int base_i = nt16_index(nt);
-    /* Minus reads: stored SEQ is 5'->3' (SAM spec) but qpos walks CIGAR order
-     * (left->right); complement the base into the reference frame (parity
-     * with countmut 0.0.x + pysam pairs). */
-    if (s == 1 && base_i < 4) base_i = 3 - base_i;
+    /* Reverse-oriented reads: stored SEQ is 5'->3' (SAM spec) but qpos walks
+     * CIGAR order (left->right); complement the base into the reference frame
+     * (parity with countmut 0.0.x + pysam pairs).  This must key on the read's
+     * own orientation (bam_is_rev), NOT on the biological strand s: for paired
+     * reads an R2 mapped forward has bio_strand=1 yet is NOT reverse-oriented,
+     * so complementing on s would wrongly flip its bases (A<->T). */
+    if (bam_is_rev(b) && base_i < 4) base_i = 3 - base_i;
     int qual = (int)bam_get_qual(b)[qpos];
     if (direct) {
         int cat = rslot;
@@ -1040,7 +1044,9 @@ static void count_interval_readwalk(worker_t *w, const cm_config *cfg, bam_hdr_t
                             if (qpos >= qlen) break;
                             uint8_t nt = bam_seqi(bam_get_seq(b), qpos);
                             int base_i = nt16_index(nt);
-                            if (s == 1 && base_i < 4) base_i = 3 - base_i;
+                            /* Complement on read orientation (bam_is_rev), not
+                             * the biological strand s (R2 differs). */
+                            if (bam_is_rev(b) && base_i < 4) base_i = 3 - base_i;
                             int cat = 0;
                             if (w->expr && cm_expr_has_read(w->expr) && !cm_expr_read_constant(w->expr)) {
                                 cat = cm_expr_route(w->expr, b, hdr->target_name[tid],
@@ -1110,7 +1116,9 @@ static void count_interval_readwalk(worker_t *w, const cm_config *cfg, bam_hdr_t
                                     int in_ovl = ((int)qpos >= olo && (int)qpos < ohi);
                                     uint8_t nt = bam_seqi(bam_get_seq(b), qpos);
                                     int base_i = nt16_index(nt);
-                                    if (s == 1 && base_i < 4) base_i = 3 - base_i;
+                                    /* Complement on read orientation (bam_is_rev),
+                                     * not the biological strand s (R2 differs). */
+                                    if (bam_is_rev(b) && base_i < 4) base_i = 3 - base_i;
                                     int cat = 0;
                                     if (w->expr && cm_expr_has_read(w->expr) && !cm_expr_read_constant(w->expr)) {
                                         cat = cm_expr_route(w->expr, b, hdr->target_name[tid],

{countmut-0.3.1 → countmut-0.3.2/countmut.egg-info}/PKG-INFO

@@ -1,6 +1,6 @@
 Metadata-Version: 2.4
 Name: countmut
-Version: 0.3.1
+Version: 0.3.2
 Summary: Unified ultra-fast strand-aware mutation counter (C backend + samtools-style -e/-p filters)
 Author-email: Ye Chang <yech1990@gmail.com>
 License-Expression: MIT

{countmut-0.3.1 → countmut-0.3.2}/pyproject.toml

@@ -4,7 +4,7 @@ build-backend = "setuptools.build_meta"
 
 [project]
 name = "countmut"
-version = "0.3.1"
+version = "0.3.2"
 description = "Unified ultra-fast strand-aware mutation counter (C backend + samtools-style -e/-p filters)"
 readme = "README.md"
 license = "MIT"

{countmut-0.3.1 → countmut-0.3.2}/tests/test_correctness.py

@@ -91,6 +91,40 @@ def indel_data(tmp_path_factory):
     )
 
 
+# ---------------------------------------------------------------------------
+# BUG: base complement keyed on biological strand (s) instead of read
+# orientation (bam_is_rev).  For paired data an R2 read mapped forward has
+# bio_strand=1 yet is NOT reverse-oriented, so its bases were wrongly
+# reverse-complemented (A<->T), turning a pure-reference site into a 50/50
+# A/T mix.  Regression: R2-forward reads must count their stored bases as-is.
+# ---------------------------------------------------------------------------
+def test_r2_forward_not_complemented(tmp_path):
+    root = str(tmp_path / "r2f")
+    os.makedirs(root, exist_ok=True)
+    # reference base at 0-based 0 is 'A'.  An R2 read mapped FORWARD stores the
+    # base as-is (A), so a pure-A site must stay A, not flip to T.
+    bam, fa = _write_bam(
+        root,
+        [
+            # R1 forward (flag 99) and R2 forward (flag 35) covering 0-based 0.
+            lambda h: _mk(h, "frag1", 99, 0, "ACGTACGTAC"),
+            lambda h: _mk(h, "frag1", 35, 0, "ACGTACGTAC"),
+            # R1 reverse (flag 83) and R2 reverse (flag 147): stored SEQ is the
+            # reverse complement, so they must be complemented back to A.
+            lambda h: _mk(h, "frag2", 83, 0, "ACGTACGTAC"),
+            lambda h: _mk(h, "frag2", 147, 0, "ACGTACGTAC"),
+        ],
+    )
+    _h, rows = run_c(bam, fa, engine="read-walk", region="chr1:1-5")
+    # rows: [chrom, pos, ref, depth, a, c, g, t, n]  (strandless default)
+    row = [r for r in rows if r[1] == 1][0]
+    a, c, g, t = row[4], row[5], row[6], row[7]
+    # All four reads carry reference 'A' at 0-based 0 -> A must dominate.
+    assert a > 0 and a >= c + g + t, (row)
+    # The bug produced ~50/50 A/T; assert T is a small minority.
+    assert t <= a * 0.1, (row)
+
+
 # ---------------------------------------------------------------------------
 # BUG: '-' mutation-row motif reverse-complemented with reference-forward bases
 # ---------------------------------------------------------------------------