Add MZ:i tag as a check for base modification validity. (#1590)

If a sequence is hard-clipped after calling the base modifications,
then the tool may, or may not, update the MM and ML tags accordingly.
We have no way of distinguishing these two cases.  While the base
modification parsing code already detects overflows where the
coordinates go beyond the sequence end, this isn't fool proof,
especially if the clipping is short.

So instead we have an (as yet unwritten) proposal of MZ:i tag holding
the sequence length, to be written at the same time as the MM and ML
tags.  This can then be used as a sanity check later on, to detect
cases where the sequence has changed length via a tool that is unaware
of base modifications.

TODO: as a separate PR, we should add a new API that can trim bases
off the start/end of MM/ML strings to make it trivial for tools that
are doing hard clipping via htslib.  (Indeed we don't even have an API
for SEQ/QUAL either, so it can do all together).  This would make it
far easier for people to keep everything in sync, and this code could
then also update MZ while it's at it.  That's new API though so it can
arrive as a separate commit.

See samtools/hts-specs#646

sam.c

@@ -6221,14 +6221,24 @@ int bam_parse_basemod(const bam1_t *b, hts_base_mod_state *state) {
     if (!mm)
         return 0;
     if (mm[0] != 'Z') {
-        hts_log_error("MM tag is not of type Z");
+        hts_log_error("%s: MM tag is not of type Z", bam_get_qname(b));
+        return -1;
+    }
+
+    uint8_t *mi = bam_aux_get(b, "MZ");
+    if (mi && bam_aux2i(mi) != b->core.l_qseq) {
+        // bam_aux2i with set errno = EINVAL and return 0 if the tag
+        // isn't integer, but 0 will be a seq-length mismatch anyway so
+        // triggers an error here too.
+        hts_log_error("%s: MM/MZ data length is incompatible with"
+                      " SEQ length", bam_get_qname(b));
         return -1;
     }
 
     uint8_t *ml = bam_aux_get(b, "ML");
     if (!ml) ml = bam_aux_get(b, "Ml");
     if (ml && (ml[0] != 'B' || ml[1] != 'C')) {
-        hts_log_error("ML tag is not of type B,C");
+        hts_log_error("%s: ML tag is not of type B,C", bam_get_qname(b));
         return -1;
     }
     uint8_t *ml_end = ml ? ml+6 + le_to_u32(ml+2) : NULL;
@@ -6314,7 +6324,8 @@ int bam_parse_basemod(const bam1_t *b, hts_base_mod_state *state) {
 
                     delta = strtol(cp, &cp_end, 10);
                     if (cp_end == cp) {
-                        hts_log_error("Hit end of MM tag. Missing semicolon?");
+                        hts_log_error("%s: Hit end of MM tag. Missing "
+                                      "semicolon?", bam_get_qname(b));
                         return -1;
                     }
 
@@ -6343,8 +6354,8 @@ int bam_parse_basemod(const bam1_t *b, hts_base_mod_state *state) {
                 state->implicit [mod_num] = implicit;
 
                 if (delta < 0) {
-                    hts_log_error("MM tag refers to bases beyond sequence "
-                                  "length");
+                    hts_log_error("%s: MM tag refers to bases beyond sequence "
+                                  "length", bam_get_qname(b));
                     return -1;
                 }
                 state->MMcount  [mod_num] = delta;
@@ -6359,7 +6370,8 @@ int bam_parse_basemod(const bam1_t *b, hts_base_mod_state *state) {
                 }
 
                 if (++mod_num >= MAX_BASE_MOD) {
-                    hts_log_error("Too many base modification types");
+                    hts_log_error("%s: Too many base modification types",
+                                  bam_get_qname(b));
                     return -1;
                 }
                 ms++; n++;
@@ -6377,7 +6389,8 @@ int bam_parse_basemod(const bam1_t *b, hts_base_mod_state *state) {
                     }
                 }
                 if (ml > ml_end) {
-                    hts_log_error("Insufficient number of entries in ML tag");
+                    hts_log_error("%s: Insufficient number of entries in ML "
+                                  "tag", bam_get_qname(b));
                     return -1;
                 }
             } else {
@@ -6389,7 +6402,8 @@ int bam_parse_basemod(const bam1_t *b, hts_base_mod_state *state) {
                         cp++;
             }
             if (!*cp) {
-                hts_log_error("Hit end of MM tag. Missing semicolon?");
+                hts_log_error("%s: Hit end of MM tag. Missing semicolon?",
+                              bam_get_qname(b));
                 return -1;
             }
         }

test/base_mods/MM-MZf1.sam

@@ -0,0 +1,5 @@
+@SQ	SN:I	LN:999
+r1	0	I	1	0	36M	*	0	0	AGCTCTCCAGAGTCGNACGCCATYCGCGCGCCACCA	DF?GCH88.EG8.7@E9G8A?H9.:C?8,@,,9F@A	Mm:Z:C+m,2,2,1,4,1;C+h,6,7;N+n,15,2;	Ml:B:C,128,153,179,204,230,159,6,215,240	MZ:i:37
+r1-	16	I	1	0	36M	*	0	0	AGCTCTCCAGAGTCGNACGCCATYCGCGCGCCACCA	DF?GCH88.EG8.7@E9G8A?H9.:C?8,@,,9F@A	Mm:Z:G-m,0,1,4,1,2;G-h,0,7;N-n,17,2;	Ml:B:C,230,204,179,153,128,6,159,240,215
+r2	0	I	4	0	3S33M	*	0	0	AGCTCTCCAGAGTCGNACGCCATYCGCGCGCCACCA	DF?GCH88.EG8.7@E9G8A?H9.:C?8,@,,9F@A	Mm:Z:C+m,2,2,1,4,1;C+h,6,7;N+n,15,2;	Ml:B:C,128,153,179,204,230,159,6,215,240
+r3	0	I	11	0	10S20M6S	*	0	0	AGCTCTCCAGAGTCGNACGCCATYCGCGCGCCACCA	DF?GCH88.EG8.7@E9G8A?H9.:C?8,@,,9F@A	Mm:Z:C+mh,2,2,0,0,4,1;N+n,15,2;	Ml:B:C,128,0,153,0,0,159,179,0,204,0,230,6,215,240	MZ:i:36

test/base_mods/MM-MZf2.sam

@@ -0,0 +1,5 @@
+@SQ	SN:I	LN:999
+r1	0	I	1	0	36M	*	0	0	AGCTCTCCAGAGTCGNACGCCATYCGCGCGCCACCA	DF?GCH88.EG8.7@E9G8A?H9.:C?8,@,,9F@A	Mm:Z:C+m,2,2,1,4,1;C+h,6,7;N+n,15,2;	Ml:B:C,128,153,179,204,230,159,6,215,240	MZ:i:36
+r1-	16	I	1	0	36M	*	0	0	AGCTCTCCAGAGTCGNACGCCATYCGCGCGCCACCA	DF?GCH88.EG8.7@E9G8A?H9.:C?8,@,,9F@A	Mm:Z:G-m,0,1,4,1,2;G-h,0,7;N-n,17,2;	Ml:B:C,230,204,179,153,128,6,159,240,215
+r2	0	I	4	0	3S33M	*	0	0	AGCTCTCCAGAGTCGNACGCCATYCGCGCGCCACCA	DF?GCH88.EG8.7@E9G8A?H9.:C?8,@,,9F@A	Mm:Z:C+m,2,2,1,4,1;C+h,6,7;N+n,15,2;	Ml:B:C,128,153,179,204,230,159,6,215,240
+r3	0	I	11	0	10S20M6S	*	0	0	AGCTCTCCAGAGTCGNACGCCATYCGCGCGCCACCA	DF?GCH88.EG8.7@E9G8A?H9.:C?8,@,,9F@A	Mm:Z:C+mh,2,2,0,0,4,1;N+n,15,2;	Ml:B:C,128,0,153,0,0,159,179,0,204,0,230,6,215,240	MZ:f:36

test/base_mods/MM-MZp.sam

@@ -0,0 +1,5 @@
+@SQ	SN:I	LN:999
+r1	0	I	1	0	36M	*	0	0	AGCTCTCCAGAGTCGNACGCCATYCGCGCGCCACCA	DF?GCH88.EG8.7@E9G8A?H9.:C?8,@,,9F@A	Mm:Z:C+m,2,2,1,4,1;C+h,6,7;N+n,15,2;	Ml:B:C,128,153,179,204,230,159,6,215,240	MZ:i:36
+r1-	16	I	1	0	36M	*	0	0	AGCTCTCCAGAGTCGNACGCCATYCGCGCGCCACCA	DF?GCH88.EG8.7@E9G8A?H9.:C?8,@,,9F@A	Mm:Z:G-m,0,1,4,1,2;G-h,0,7;N-n,17,2;	Ml:B:C,230,204,179,153,128,6,159,240,215
+r2	0	I	4	0	3S33M	*	0	0	AGCTCTCCAGAGTCGNACGCCATYCGCGCGCCACCA	DF?GCH88.EG8.7@E9G8A?H9.:C?8,@,,9F@A	Mm:Z:C+m,2,2,1,4,1;C+h,6,7;N+n,15,2;	Ml:B:C,128,153,179,204,230,159,6,215,240
+r3	0	I	11	0	10S20M6S	*	0	0	AGCTCTCCAGAGTCGNACGCCATYCGCGCGCCACCA	DF?GCH88.EG8.7@E9G8A?H9.:C?8,@,,9F@A	Mm:Z:C+mh,2,2,0,0,4,1;N+n,15,2;	Ml:B:C,128,0,153,0,0,159,179,0,204,0,230,6,215,240	MZ:i:36

test/base_mods/MM-multi.sam

@@ -3,5 +3,5 @@
 @CO	r2 has them combined together, for example as produced by
 @CO	a joint basecaller which assigns probabilities to all
 @CO	trained events simultaneously.
-r1	0	*	0	0	*	*	0	0	AGCTCTCCAGAGTCGNACGCCATYCGCGCGCCACCA	*	Mm:Z:C+m,2,2,1,4,1;C+h,6,7;N+n,15,2;	Ml:B:C,128,153,179,204,230,159,6,215,240
+r1	0	*	0	0	*	*	0	0	AGCTCTCCAGAGTCGNACGCCATYCGCGCGCCACCA	*	Mm:Z:C+m,2,2,1,4,1;C+h,6,7;N+n,15,2;	Ml:B:C,128,153,179,204,230,159,6,215,240	MZ:i:36
 r2	0	*	0	0	*	*	0	0	AGCTCTCCAGAGTCGNACGCCATYCGCGCGCCACCA	*	Mm:Z:C+mh,2,2,0,0,4,1;N+n,15;	Ml:B:C,77,159,103,133,128,108,154,82,179,57,204,31,240

test/base_mods/base-mods.sh

@@ -31,5 +31,6 @@ test_mod="../test_mod"
 pileup_mod="../pileup_mod"
 
 test_driver $@
+rm _err.tmp _out.tmp
 
 exit $?

test/base_mods/base-mods.tst

@@ -42,3 +42,9 @@ P MM-explicit-x.out  $test_mod -x MM-explicit.sam
 # Pileup testing
 P MM-pileup.out $pileup_mod < MM-pileup.sam
 P MM-pileup2.out $pileup_mod < MM-pileup2.sam
+
+# Validation testing.  We just care about exit status here, but the
+# test data is a copy of MM-pileup.sam so that suffices too.
+P MM-pileup.out $pileup_mod < MM-MZp.sam
+F MM-pileup.out $pileup_mod < MM-MZf1.sam
+F MM-pileup.out $pileup_mod < MM-MZf2.sam

test/pileup_mod.c

@@ -73,7 +73,8 @@ void process_pileup(sam_hdr_t *h, const bam_pileup1_t *p,
 // as each new read is added or removed from the pileups.
 int pileup_cd_create(void *data, const bam1_t *b, bam_pileup_cd *cd) {
     hts_base_mod_state *m = hts_base_mod_state_alloc();
-    bam_parse_basemod(b, m);
+    if (bam_parse_basemod(b, m) < 0)
+        return -1;
     cd->p = m;
     return 0;
 }
@@ -201,7 +202,7 @@ int main(int argc, char **argv) {
     bam_plp_destructor(iter, pileup_cd_destroy);
 
     const bam_pileup1_t *p;
-    int tid, pos, n;
+    int tid, pos, n = 0;
     while ((p = bam_plp_auto(iter, &tid, &pos, &n)) != 0) {
         switch (compact) {
         case 0:
@@ -221,5 +222,5 @@ int main(int argc, char **argv) {
     bam_destroy1(b);
     sam_hdr_destroy(h);
 
-    return 0;
+    return n != 0;
 }