Support for clipping reads that extend past their mate

src/main/scala/com/fulcrumgenomics/alignment/Alignment.scala

@@ -91,33 +91,71 @@ object Cigar {
 }
 
 /** A data class for holding statistics about a [[Cigar]]. */
-private[alignment] case class CigarStats(lengthOnQuery: Int, lengthOnTarget: Int, alignedBases: Int, clippedBases: Int)
+private[alignment] case class CigarStats(lengthOnQuery: Int,
+                                         lengthOnTarget: Int,
+                                         alignedBases: Int,
+                                         clippedBases: Int,
+                                         softClippedFivePrimeBases: Int,
+                                         softClippedThreePrimeBases: Int)
 
 /** Companion object for [[Cigar]]. */
 private[alignment] object CigarStats {
 
   /** Build a [[CigarStats]] from a [[Cigar]]. */
   private[alignment] def apply(cigar: Cigar): CigarStats = {
-    var lengthOnQuery  = 0
-    var lengthOnTarget = 0
-    var alignedBases   = 0
-    var clippedBases   = 0
+    var lengthOnQuery              = 0
+    var lengthOnTarget             = 0
+    var alignedBases               = 0
+    var clippedBases               = 0
+    var softClippedFivePrimeBases  = 0
+    var softClippedThreePrimeBases = 0
+    var index                      = 0
+
+    // leading clipping
+    while (index < cigar.elems.length && cigar.elems(index).operator.isClipping) {
+      val elem = cigar.elems(index)
+      lengthOnQuery  += elem.lengthOnQuery
+      lengthOnTarget += elem.lengthOnTarget
+      clippedBases += elem.length
+      if (elem.operator == CigarOperator.S) softClippedFivePrimeBases += elem.length
+      index += 1
+    }
 
-    forloop(0, cigar.elems.length) { index =>
+    // aligned bases
+    while (index < cigar.elems.length && !cigar.elems(index).operator.isClipping) {
       val elem = cigar.elems(index)
       lengthOnQuery  += elem.lengthOnQuery
       lengthOnTarget += elem.lengthOnTarget
-      if (elem.operator.isAlignment)     alignedBases += elem.length
-      else if (elem.operator.isClipping) clippedBases += elem.length
+      if (elem.operator.isAlignment) alignedBases += elem.length
+      index += 1
     }
 
+    // trailing clipping
+    while (index < cigar.elems.length && cigar.elems(index).operator.isClipping) {
+      val elem = cigar.elems(index)
+      lengthOnQuery  += elem.lengthOnQuery
+      lengthOnTarget += elem.lengthOnTarget
+      clippedBases += elem.length
+      if (elem.operator == CigarOperator.S) softClippedThreePrimeBases += elem.length
+      index += 1
+    }
+
+    require(index == cigar.elems.length,
+      s"Invalid cigar, did not consume all the elements: ${Cigar(cigar.elems.take(index))}[${Cigar(cigar.elems.drop(index))}]"
+    )
+
     CigarStats(
-      lengthOnQuery  = lengthOnQuery,
-      lengthOnTarget = lengthOnTarget,
-      alignedBases   = alignedBases,
-      clippedBases   = clippedBases
+      lengthOnQuery              = lengthOnQuery,
+      lengthOnTarget             = lengthOnTarget,
+      alignedBases               = alignedBases,
+      clippedBases               = clippedBases,
+      softClippedFivePrimeBases  = softClippedFivePrimeBases,
+      softClippedThreePrimeBases = softClippedThreePrimeBases
     )
   }
+
+  /** Build a [[CigarStats]] from a string representation of the cigar. */
+  private[alignment] def apply(cigar: String): CigarStats = this.apply(Cigar(cigar))
 }
 
 /**
@@ -163,6 +201,12 @@ case class Cigar(elems: IndexedSeq[CigarElem]) extends Iterable[CigarElem] {
   /** Returns the number of bases that are clipped between the two sequences. */
   def clippedBases: Int = stats.clippedBases
 
+  /** Returns the number of bases that are soft-clipped on the 5' end of this sequence.  Ignores hard-clips. */
+  def softClippedFivePrimeBases: Int = stats.softClippedFivePrimeBases
+
+  /** Returns the number of bases that are soft-clipped on the 3' end of this sequence.  Ignores hard-clips. */
+  def softClippedThreePrimeBases: Int = stats.softClippedThreePrimeBases
+
   /** Truncates the cigar based on either query or target length cutoff. */
   private def truncate(len: Int, shouldCount: CigarElem => Boolean): Cigar = {
     var pos = 1
@@ -297,21 +341,21 @@ case class Alignment(query: Array[Byte],
 
       cig.operator match {
         case CigarOperator.INSERTION =>
-          forloop (from=0, until=len) { i =>
+          forloop (from=0, until=len) { _ =>
             queryBuffer.append(this.query(qOffset).toChar)
             alignBuffer.append(gapChar)
             targetBuffer.append(padChar)
             qOffset += 1
           }
         case CigarOperator.DELETION =>
-          forloop (from=0, until=len) { i =>
+          forloop (from=0, until=len) { _ =>
             queryBuffer.append(padChar)
             alignBuffer.append(gapChar)
             targetBuffer.append(this.target(tOffset).toChar)
             tOffset += 1
           }
         case op =>
-          forloop (from=0, until=len) { i =>
+          forloop (from=0, until=len) { _ =>
             val q = this.query(qOffset).toChar
             val t = this.target(tOffset).toChar
             queryBuffer.append(q)

src/main/scala/com/fulcrumgenomics/bam/ClipBam.scala

@@ -78,13 +78,14 @@ class ClipBam
   @arg(          doc="Require at least this number of bases to be clipped on the 3' end of R1") val readOneThreePrime: Int = 0,
   @arg(          doc="Require at least this number of bases to be clipped on the 5' end of R2") val readTwoFivePrime: Int  = 0,
   @arg(          doc="Require at least this number of bases to be clipped on the 3' end of R2") val readTwoThreePrime: Int = 0,
-  @arg(          doc="Clip overlapping reads.") val clipOverlappingReads: Boolean = false
+  @arg(          doc="Clip overlapping reads.", mutex=Array("clipBasesPastMate")) val clipOverlappingReads: Boolean = false,
+  @arg(          doc="Clip reads who sequence past the start of their mate.", mutex=Array("clipOverlappingReads")) val clipBasesPastMate: Boolean = false
 ) extends FgBioTool with LazyLogging {
   Io.assertReadable(input)
   Io.assertReadable(ref)
   Io.assertCanWriteFile(output)
 
-  validate(upgradeClipping || clipOverlappingReads || Seq(readOneFivePrime, readOneThreePrime, readTwoFivePrime, readTwoThreePrime).exists(_ != 0),
+  validate(upgradeClipping || clipOverlappingReads || clipBasesPastMate || Seq(readOneFivePrime, readOneThreePrime, readTwoFivePrime, readTwoThreePrime).exists(_ != 0),
     "At least one clipping option is required")
 
   private val clipper = new SamRecordClipper(mode=clippingMode, autoClipAttributes=autoClipAttributes)
@@ -164,7 +165,8 @@ class ClipBam
         priorBasesClipped   = priorBasesClipped,
         numFivePrime        = numFivePrime,
         numThreePrime       = numThreePrime,
-        numOverlappingBases = 0
+        numOverlappingBases = 0,
+        numExtendingBases   = 0
       )
     }
   }
@@ -182,9 +184,13 @@ class ClipBam
     val numReadTwoThreePrime = this.clipper.clip3PrimeEndOfRead(r2, readTwoThreePrime)
 
     val (numOverlappingBasesReadOne, numOverlappingBasesReadTwo) = {
-      if (clipOverlappingReads && r1.isFrPair) {
-        if (r1.positiveStrand) clipOverlappingReads(r1, r2) else clipOverlappingReads(r2, r1)
-      } else (0, 0)
+      if (clipOverlappingReads && r1.isFrPair) this.clipper.clipOverlappingReads(r1, r2)
+      else (0, 0)
+    }
+
+    val (numExtendingPastMateStartReadOne, numExtendingPastMateStartReadTwo) = {
+      if (clipBasesPastMate && r1.isFrPair) this.clipper.clipExtendingPastMateEnds(r1, r2)
+      else (0, 0)
     }
 
     r1Metric.foreach { m =>
@@ -193,7 +199,8 @@ class ClipBam
         priorBasesClipped   = priorBasesClippedReadOne,
         numFivePrime        = numReadOneFivePrime,
         numThreePrime       = numReadOneThreePrime,
-        numOverlappingBases = numOverlappingBasesReadOne
+        numOverlappingBases = numOverlappingBasesReadOne,
+        numExtendingBases   = numExtendingPastMateStartReadOne
       )
     }
 
@@ -203,27 +210,11 @@ class ClipBam
         priorBasesClipped   = priorBasesClippedReadTwo,
         numFivePrime        = numReadTwoFivePrime,
         numThreePrime       = numReadTwoThreePrime,
-        numOverlappingBases = numOverlappingBasesReadTwo
+        numOverlappingBases = numOverlappingBasesReadTwo,
+        numExtendingBases   = numExtendingPastMateStartReadTwo
       )
     }
   }
-
-  /** Clips overlapping reads, where both ends of the read pair are mapped to the same chromosome, and in FR orientation. */
-  protected def clipOverlappingReads(f: SamRecord, r: SamRecord): (Int, Int) = {
-    var numOverlappingBasesReadOne: Int = 0
-    var numOverlappingBasesReadTwo: Int = 0
-    // What we really want is to trim by the number of _reference_ bases not read bases,
-    // in order to eliminate overlap.  We could do something very complicated here, or
-    // we could just trim read bases in a loop until the overlap is eliminated!
-    while (f.end >= r.start && f.mapped && r.mapped) {
-      val lengthToClip = f.end - r.start + 1
-      val firstHalf    = lengthToClip / 2
-      val secondHalf   = lengthToClip - firstHalf // safe guard against rounding on odd lengths
-      numOverlappingBasesReadOne += this.clipper.clip3PrimeEndOfAlignment(f, firstHalf)
-      numOverlappingBasesReadTwo += this.clipper.clip3PrimeEndOfAlignment(r, secondHalf)
-    }
-    (numOverlappingBasesReadOne, numOverlappingBasesReadTwo)
-  }
 }
 
 sealed trait ReadType extends EnumEntry
@@ -246,13 +237,15 @@ object ReadType extends FgBioEnum[ReadType] {
   * @param reads_clipped_five_prime The number of reads with the 5' end clipped.
   * @param reads_clipped_three_prime The number of reads with the 3' end clipped.
   * @param reads_clipped_overlapping The number of reads clipped due to overlapping reads.
+  * @param reads_clipped_extending The number of reads clipped due to a read extending past its mate.
   * @param reads_unmapped The number of reads that became unmapped due to clipping.
   * @param bases The number of aligned bases after clipping.
   * @param bases_clipped_pre The number of bases clipped prior to clipping with [[ClipBam]].
   * @param bases_clipped_post The number of bases clipped after clipping with [[ClipBam]], including bases from reads that became unmapped.
   * @param bases_clipped_five_prime The number of bases clipped on the 5' end of the read.
   * @param bases_clipped_three_prime The number of bases clipped on the 3 end of the read.
   * @param bases_clipped_overlapping The number of bases clipped due to overlapping reads.
+  * @param bases_clipped_extending The number of bases clipped due to a read extending past its mate.
   */
 case class ClippingMetrics
 (read_type: ReadType,
@@ -263,14 +256,16 @@ case class ClippingMetrics
  var reads_clipped_five_prime: Long = 0,
  var reads_clipped_three_prime: Long = 0,
  var reads_clipped_overlapping: Long = 0,
+ var reads_clipped_extending: Long = 0,
  var bases: Long = 0,
  var bases_clipped_pre: Long = 0,
  var bases_clipped_post: Long = 0,
  var bases_clipped_five_prime: Long = 0,
  var bases_clipped_three_prime: Long = 0,
- var bases_clipped_overlapping: Long = 0
+ var bases_clipped_overlapping: Long = 0,
+ var bases_clipped_extending: Long = 0,
 ) extends Metric {
-  def update(rec: SamRecord, priorBasesClipped: Int, numFivePrime: Int, numThreePrime: Int, numOverlappingBases: Int): Unit = {
+  def update(rec: SamRecord, priorBasesClipped: Int, numFivePrime: Int, numThreePrime: Int, numOverlappingBases: Int, numExtendingBases: Int): Unit = {
     this.reads                      += 1
     this.bases                      += rec.cigar.alignedBases
     if (priorBasesClipped > 0) {
@@ -289,11 +284,15 @@ case class ClippingMetrics
       this.reads_clipped_overlapping += 1
       this.bases_clipped_overlapping += numOverlappingBases
     }
-    val additionalClippedBases = numFivePrime + numThreePrime + numOverlappingBases
-    val totalClippedBasees = additionalClippedBases + priorBasesClipped
-    if (totalClippedBasees > 0) {
+    if (numExtendingBases > 0) {
+      this.reads_clipped_extending += 1
+      this.bases_clipped_extending += numExtendingBases
+    }
+    val additionalClippedBases = numFivePrime + numThreePrime + numOverlappingBases + numExtendingBases
+    val totalClippedBases = additionalClippedBases + priorBasesClipped
+    if (totalClippedBases > 0) {
       this.reads_clipped_post        += 1
-      this.bases_clipped_post        += totalClippedBasees
+      this.bases_clipped_post        += totalClippedBases
       if (rec.unmapped && additionalClippedBases > 0) this.reads_unmapped += 1
     }
   }
@@ -306,11 +305,13 @@ case class ClippingMetrics
     this.reads_clipped_five_prime  += metric.sumBy(_.reads_clipped_five_prime)
     this.reads_clipped_three_prime += metric.sumBy(_.reads_clipped_three_prime)
     this.reads_clipped_overlapping += metric.sumBy(_.reads_clipped_overlapping)
+    this.reads_clipped_extending   += metric.sumBy(_.reads_clipped_extending)
     this.bases                     += metric.sumBy(_.bases)
     this.bases_clipped_pre         += metric.sumBy(_.bases_clipped_pre)
     this.bases_clipped_post        += metric.sumBy(_.bases_clipped_post)
     this.bases_clipped_five_prime  += metric.sumBy(_.bases_clipped_five_prime)
     this.bases_clipped_three_prime += metric.sumBy(_.bases_clipped_three_prime)
     this.bases_clipped_overlapping += metric.sumBy(_.bases_clipped_overlapping)
+    this.bases_clipped_extending   += metric.sumBy(_.bases_clipped_extending)
   }
 }