diff --git a/biopet-framework/src/main/scala/nl/lumc/sasc/biopet/tools/ExtractAlignedFastq.scala b/biopet-framework/src/main/scala/nl/lumc/sasc/biopet/tools/ExtractAlignedFastq.scala
index cdbc13e996e55e200d322bf9cc2691e09834fa5f..83a607de4a035013cbf82072223f533380ff942c 100644
--- a/biopet-framework/src/main/scala/nl/lumc/sasc/biopet/tools/ExtractAlignedFastq.scala
+++ b/biopet-framework/src/main/scala/nl/lumc/sasc/biopet/tools/ExtractAlignedFastq.scala
@@ -9,19 +9,20 @@ import java.io.File
import scala.collection.mutable.{ Set => MSet }
import scala.collection.JavaConverters._
-import htsjdk.samtools.SAMFileReader
import htsjdk.samtools.QueryInterval
+import htsjdk.samtools.SamReaderFactory
+import htsjdk.samtools.ValidationStringency
import htsjdk.samtools.fastq.{ BasicFastqWriter, FastqReader, FastqRecord }
-import htsjdk.tribble.Feature
-import htsjdk.tribble.BasicFeature
+import htsjdk.samtools.util.Interval
import nl.lumc.sasc.biopet.core.ToolCommand
object ExtractAlignedFastq extends ToolCommand {
type FastqPair = (FastqRecord, FastqRecord)
+
/**
- * Function to create iterator over features given input interval string
+ * Function to create iterator over Interval given input interval string
*
* Valid interval strings are either of these:
* - chr5:10000-11000
@@ -38,7 +39,7 @@ object ExtractAlignedFastq extends ToolCommand {
*
* @param inStrings iterable yielding input interval string
*/
- def makeFeatureFromString(inStrings: Iterable[String]): Iterator[Feature] = {
+ def makeIntervalFromString(inStrings: Iterable[String]): Iterator[Interval] = {
// FIXME: can we combine these two patterns into one regex?
// matches intervals with start and end coordinates
@@ -47,15 +48,16 @@ object ExtractAlignedFastq extends ToolCommand {
val ptn2 = """([\w_-]+):([\d.,]+)""".r
// make ints from coordinate strings
// NOTE: while it is possible for coordinates to exceed Int.MaxValue, we are limited
- // by the BasicFeature constructor only accepting ints
+ // by the Interval constructor only accepting ints
def intFromCoord(s: String): Int = s.replaceAll(",", "").replaceAll("\\.", "").toInt
inStrings.map(x => x match {
- case ptn1(chr, start, end) => new BasicFeature(chr, intFromCoord(start), intFromCoord(end))
- case ptn2(chr, start) => val startCoord = intFromCoord(start)
- new BasicFeature(chr, startCoord, startCoord)
- case _ => throw new IllegalArgumentException("Invalid interval string: " + x)
- })
+ case ptn1(chr, start, end) => new Interval(chr, intFromCoord(start), intFromCoord(end))
+ case ptn2(chr, start) =>
+ val startCoord = intFromCoord(start)
+ new Interval(chr, startCoord, startCoord)
+ case _ => throw new IllegalArgumentException("Invalid interval string: " + x)
+ })
.toIterator
}
@@ -69,34 +71,30 @@ object ExtractAlignedFastq extends ToolCommand {
* @param commonSuffixLength length of suffix common to all read pairs
* @return
*/
- def makeMembershipFunction(iv: Iterator[Feature],
+ def makeMembershipFunction(iv: Iterator[Interval],
inAln: File,
minMapQ: Int = 0,
- commonSuffixLength: Int = 0
- ): (FastqPair => Boolean) = {
-
- /** function to make interval queries for BAM files */
- def makeQueryInterval(aln: SAMFileReader, feat: Feature): QueryInterval =
- if (aln.getFileHeader.getSequenceIndex(feat.getChr) > -1)
- aln.makeQueryInterval(feat.getChr, feat.getStart, feat.getEnd)
- else if (feat.getChr.startsWith("chr")
- && aln.getFileHeader.getSequenceIndex(feat.getChr.substring(3)) > -1)
- aln.makeQueryInterval(feat.getChr.substring(3), feat.getStart, feat.getEnd)
- else if (!feat.getChr.startsWith("chr")
- && aln.getFileHeader.getSequenceIndex("chr" + feat.getChr) > -1)
- aln.makeQueryInterval("chr" + feat.getChr, feat.getStart, feat.getEnd)
- else
- throw new IllegalStateException("Unexpected feature: " + feat.toString)
+ commonSuffixLength: Int = 0): (FastqPair => Boolean) = {
- val inAlnReader = new SAMFileReader(inAln)
+ val inAlnReader = SamReaderFactory
+ .make()
+ .validationStringency(ValidationStringency.LENIENT)
+ .open(inAln)
require(inAlnReader.hasIndex)
+ def getSequenceIndex(name: String): Int = inAlnReader.getFileHeader.getSequenceIndex(name) match {
+ case x if x >= 0 =>
+ x
+ case otherwise =>
+ throw new IllegalArgumentException("Chromosome " + name + " is not found in the alignment file")
+ }
+
val queries: Array[QueryInterval] = iv.toList
- // sort features
- .sortBy(x => (x.getChr, x.getStart, x.getEnd))
- // turn them into QueryInterval objects
- .map(x => makeQueryInterval(inAlnReader, x))
- // return as an array
+ // sort Interval
+ .sortBy(x => (x.getSequence, x.getStart, x.getEnd))
+ // transform to QueryInterval
+ .map(x => new QueryInterval(getSequenceIndex(x.getSequence), x.getStart, x.getEnd))
+ // cast to array
.toArray
lazy val selected: MSet[String] = inAlnReader
@@ -112,11 +110,11 @@ object ExtractAlignedFastq extends ToolCommand {
logger.debug("Adding " + x.getReadName + " to set ...")
acc += x.getReadName
}
- )
+ )
(pair: FastqPair) => pair._2 match {
- case null => selected.contains(pair._1.getReadHeader)
- case otherwise =>
+ case null => selected.contains(pair._1.getReadHeader)
+ case otherwise =>
require(commonSuffixLength < pair._1.getReadHeader.length)
require(commonSuffixLength < pair._2.getReadHeader.length)
selected.contains(pair._1.getReadHeader.dropRight(commonSuffixLength))
@@ -131,8 +129,8 @@ object ExtractAlignedFastq extends ToolCommand {
val i1 = inputFastq1.iterator.asScala
val i2 = inputFastq2 match {
- case null => Iterator.continually(null)
- case otherwise => otherwise.iterator.asScala
+ case null => Iterator.continually(null)
+ case otherwise => otherwise.iterator.asScala
}
val o1 = outputFastq1
val o2 = (inputFastq2, outputFastq2) match {
@@ -152,18 +150,18 @@ object ExtractAlignedFastq extends ToolCommand {
case (rec1, rec2) =>
o1.write(rec1)
o2.write(rec2)
- }
+ }
}
- case class Args (inputBam: File = null,
- intervals: List[String] = List.empty[String],
- inputFastq1: File = null,
- inputFastq2: File = null,
- outputFastq1: File = null,
- outputFastq2: File = null,
- minMapQ: Int = 0,
- commonSuffixLength: Int = 0) extends AbstractArgs
+ case class Args(inputBam: File = null,
+ intervals: List[String] = List.empty[String],
+ inputFastq1: File = null,
+ inputFastq2: File = null,
+ outputFastq1: File = null,
+ outputFastq2: File = null,
+ minMapQ: Int = 0,
+ commonSuffixLength: Int = 0) extends AbstractArgs
class OptParser extends AbstractOptParser {
@@ -172,36 +170,44 @@ object ExtractAlignedFastq extends ToolCommand {
|$commandName - Select aligned FASTQ records
""".stripMargin)
- opt[File]('I', "input_file") required() valueName "<bam>" action { (x, c) =>
- c.copy(inputBam = x) } validate {
+ opt[File]('I', "input_file") required () valueName "<bam>" action { (x, c) =>
+ c.copy(inputBam = x)
+ } validate {
x => if (x.exists) success else failure("Input BAM file not found")
} text "Input BAM file"
- opt[String]('r', "interval") required() unbounded() valueName "<interval>" action { (x, c) =>
+ opt[String]('r', "interval") required () unbounded () valueName "<interval>" action { (x, c) =>
// yes, we are appending and yes it's O(n) ~ preserving order is more important than speed here
- c.copy(intervals = c.intervals :+ x) } text "Interval strings"
+ c.copy(intervals = c.intervals :+ x)
+ } text "Interval strings"
- opt[File]('i', "in1") required() valueName "<fastq>" action { (x, c) =>
- c.copy(inputFastq1 = x) } validate {
+ opt[File]('i', "in1") required () valueName "<fastq>" action { (x, c) =>
+ c.copy(inputFastq1 = x)
+ } validate {
x => if (x.exists) success else failure("Input FASTQ file 1 not found")
} text "Input FASTQ file 1"
- opt[File]('j', "in2") optional() valueName "<fastq>" action { (x, c) =>
- c.copy(inputFastq1 = x) } validate {
+ opt[File]('j', "in2") optional () valueName "<fastq>" action { (x, c) =>
+ c.copy(inputFastq1 = x)
+ } validate {
x => if (x.exists) success else failure("Input FASTQ file 2 not found")
} text "Input FASTQ file 2 (default: none)"
- opt[File]('o', "out1") required() valueName "<fastq>" action { (x, c) =>
- c.copy(outputFastq1 = x) } text "Output FASTQ file 1"
+ opt[File]('o', "out1") required () valueName "<fastq>" action { (x, c) =>
+ c.copy(outputFastq1 = x)
+ } text "Output FASTQ file 1"
- opt[File]('p', "out2") optional() valueName "<fastq>" action { (x, c) =>
- c.copy(outputFastq1 = x) } text "Output FASTQ file 2 (default: none)"
+ opt[File]('p', "out2") optional () valueName "<fastq>" action { (x, c) =>
+ c.copy(outputFastq1 = x)
+ } text "Output FASTQ file 2 (default: none)"
- opt[Int]('Q', "min_mapq") optional() action { (x, c) =>
- c.copy(minMapQ = x) } text "Minimum MAPQ of reads in target region to remove (default: 0)"
+ opt[Int]('Q', "min_mapq") optional () action { (x, c) =>
+ c.copy(minMapQ = x)
+ } text "Minimum MAPQ of reads in target region to remove (default: 0)"
- opt[Int]('s', "read_suffix_length") optional() action { (x, c) =>
- c.copy(commonSuffixLength = x) } text "Length of common suffix from each read pair (default: 0)"
+ opt[Int]('s', "read_suffix_length") optional () action { (x, c) =>
+ c.copy(commonSuffixLength = x)
+ } text "Length of common suffix from each read pair (default: 0)"
note(
"""
@@ -229,7 +235,7 @@ object ExtractAlignedFastq extends ToolCommand {
.getOrElse(sys.exit(1))
val memFunc = makeMembershipFunction(
- iv = makeFeatureFromString(commandArgs.intervals),
+ iv = makeIntervalFromString(commandArgs.intervals),
inAln = commandArgs.inputBam,
minMapQ = commandArgs.minMapQ,
commonSuffixLength = commandArgs.commonSuffixLength)
diff --git a/biopet-framework/src/test/scala/nl/lumc/sasc/biopet/tools/ExtractAlignedFastqUnitTest.scala b/biopet-framework/src/test/scala/nl/lumc/sasc/biopet/tools/ExtractAlignedFastqUnitTest.scala
index d166ad4f716139fa4e1c29cc004f6f2938611b72..0dac57875f5ae3966b009d9605a2e7c7d55b5d96 100644
--- a/biopet-framework/src/test/scala/nl/lumc/sasc/biopet/tools/ExtractAlignedFastqUnitTest.scala
+++ b/biopet-framework/src/test/scala/nl/lumc/sasc/biopet/tools/ExtractAlignedFastqUnitTest.scala
@@ -14,8 +14,8 @@ import org.scalatest.mock.MockitoSugar
import org.scalatest.testng.TestNGSuite
import org.testng.annotations.{ DataProvider, Test }
+import htsjdk.samtools.util.Interval
import htsjdk.samtools.fastq.{ BasicFastqWriter, FastqReader, FastqRecord }
-import htsjdk.tribble._
class ExtractAlignedFastqUnitTest extends TestNGSuite with MockitoSugar with Matchers {
@@ -24,8 +24,8 @@ class ExtractAlignedFastqUnitTest extends TestNGSuite with MockitoSugar with Mat
private def resourceFile(p: String): File =
new File(Paths.get(getClass.getResource(p).toURI).toString)
- private def makeFeature(chr: String, start: Int, end: Int): Feature =
- new BasicFeature(chr, start ,end)
+ private def makeInterval(chr: String, start: Int, end: Int): Interval =
+ new Interval(chr, start, end)
private def makeRecord(header: String): FastqRecord =
new FastqRecord(header, "ATGC", "", "HIHI")
@@ -40,41 +40,52 @@ class ExtractAlignedFastqUnitTest extends TestNGSuite with MockitoSugar with Mat
tName + " on " + f.getName + ", read " + rName + ": "
@Test def testIntervalStartEnd() = {
- val obs = makeFeatureFromString(List("chr5:1000-1100")).next()
- val exp = new BasicFeature("chr5", 1000, 1100)
- obs.getChr should === (exp.getChr)
+ val obs = makeIntervalFromString(List("chr5:1000-1100")).next()
+ val exp = new Interval("chr5", 1000, 1100)
+ obs.getSequence should === (exp.getSequence)
obs.getStart should === (exp.getStart)
obs.getEnd should === (exp.getEnd)
}
@Test def testIntervalStartEndComma() = {
- val obs = makeFeatureFromString(List("chr5:1,000-1,100")).next()
- val exp = new BasicFeature("chr5", 1000, 1100)
- obs.getChr should === (exp.getChr)
+ val obs = makeIntervalFromString(List("chr5:1,000-1,100")).next()
+ val exp = new Interval("chr5", 1000, 1100)
+ obs.getSequence should === (exp.getSequence)
obs.getStart should === (exp.getStart)
obs.getEnd should === (exp.getEnd)
}
@Test def testIntervalStartEndDot() = {
- val obs = makeFeatureFromString(List("chr5:1.000-1.100")).next()
- val exp = new BasicFeature("chr5", 1000, 1100)
- obs.getChr should === (exp.getChr)
+ val obs = makeIntervalFromString(List("chr5:1.000-1.100")).next()
+ val exp = new Interval("chr5", 1000, 1100)
+ obs.getSequence should === (exp.getSequence)
obs.getStart should === (exp.getStart)
obs.getEnd should === (exp.getEnd)
}
@Test def testIntervalStart() = {
- val obs = makeFeatureFromString(List("chr5:1000")).next()
- val exp = new BasicFeature("chr5", 1000, 1000)
- obs.getChr should === (exp.getChr)
+ val obs = makeIntervalFromString(List("chr5:1000")).next()
+ val exp = new Interval("chr5", 1000, 1000)
+ obs.getSequence should === (exp.getSequence)
obs.getStart should === (exp.getStart)
obs.getEnd should === (exp.getEnd)
}
- @Test def testIntervalError() =
- intercept[IllegalArgumentException] {
- makeFeatureFromString(List("chr5:1000-")).next()
+ @Test def testIntervalError() = {
+ val thrown = intercept[IllegalArgumentException] {
+ makeIntervalFromString(List("chr5:1000-")).next()
+ }
+ thrown.getMessage should === ("Invalid interval string: chr5:1000-")
+ }
+
+ @Test def testMemFuncIntervalError() = {
+ val iv = Iterator(new Interval("chrP", 1, 1000))
+ val inAln = resourceFile("/single01.bam")
+ val thrown = intercept[IllegalArgumentException] {
+ makeMembershipFunction(iv, inAln)
}
+ thrown.getMessage should === ("Chromosome chrP is not found in the alignment file")
+ }
@DataProvider(name = "singleAlnProvider1", parallel = true)
def singleAlnProvider1() = {
@@ -84,20 +95,20 @@ class ExtractAlignedFastqUnitTest extends TestNGSuite with MockitoSugar with Mat
Array(
Array("adjacent left",
- makeFeature("chrQ", 30, 49), sBam01, sFastq1, sFastq1Default),
+ makeInterval("chrQ", 30, 49), sBam01, sFastq1, sFastq1Default),
Array("adjacent right",
- makeFeature("chrQ", 200, 210), sBam01, sFastq1, sFastq1Default),
+ makeInterval("chrQ", 200, 210), sBam01, sFastq1, sFastq1Default),
Array("no overlap",
- makeFeature("chrQ", 220, 230), sBam01, sFastq1, sFastq1Default),
+ makeInterval("chrQ", 220, 230), sBam01, sFastq1, sFastq1Default),
Array("partial overlap",
- makeFeature("chrQ", 430, 460), sBam01, sFastq1, sFastq1Default.updated("r04", true)),
+ makeInterval("chrQ", 430, 460), sBam01, sFastq1, sFastq1Default.updated("r04", true)),
Array("enveloped",
- makeFeature("chrQ", 693, 698), sBam01, sFastq1, sFastq1Default.updated("r03", true))
+ makeInterval("chrQ", 693, 698), sBam01, sFastq1, sFastq1Default.updated("r03", true))
)
}
@Test(dataProvider = "singleAlnProvider1")
- def testSingleBamDefault(name: String, feat: Feature, inAln: File,
+ def testSingleBamDefault(name: String, feat: Interval, inAln: File,
fastqMap: Map[String, FastqPair], resultMap: Map[String, Boolean]) = {
require(resultMap.keySet == fastqMap.keySet)
val memFunc = makeMembershipFunction(Iterator(feat), inAln)
@@ -116,16 +127,16 @@ class ExtractAlignedFastqUnitTest extends TestNGSuite with MockitoSugar with Mat
Array(
Array("less than minimum MAPQ",
- makeFeature("chrQ", 830, 890), sBam02, 60, sFastq2, sFastq2Default),
+ makeInterval("chrQ", 830, 890), sBam02, 60, sFastq2, sFastq2Default),
Array("greater than minimum MAPQ",
- makeFeature("chrQ", 830, 890), sBam02, 20, sFastq2, sFastq2Default.updated("r07", true)),
+ makeInterval("chrQ", 830, 890), sBam02, 20, sFastq2, sFastq2Default.updated("r07", true)),
Array("equal to minimum MAPQ",
- makeFeature("chrQ", 260, 320), sBam02, 30, sFastq2, sFastq2Default.updated("r01", true))
+ makeInterval("chrQ", 260, 320), sBam02, 30, sFastq2, sFastq2Default.updated("r01", true))
)
}
@Test(dataProvider = "singleAlnProvider2")
- def testSingleBamMinMapQ(name: String, feat: Feature, inAln: File, minMapQ: Int,
+ def testSingleBamMinMapQ(name: String, feat: Interval, inAln: File, minMapQ: Int,
fastqMap: Map[String, FastqPair], resultMap: Map[String, Boolean]) = {
require(resultMap.keySet == fastqMap.keySet)
val memFunc = makeMembershipFunction(Iterator(feat), inAln, minMapQ)
@@ -148,22 +159,22 @@ class ExtractAlignedFastqUnitTest extends TestNGSuite with MockitoSugar with Mat
Array(
Array("adjacent left",
- makeFeature("chrQ", 30, 49), pBam01, pFastq1, pFastq1Default),
+ makeInterval("chrQ", 30, 49), pBam01, pFastq1, pFastq1Default),
Array("adjacent right",
- makeFeature("chrQ", 200, 210), pBam01, pFastq1, pFastq1Default),
+ makeInterval("chrQ", 200, 210), pBam01, pFastq1, pFastq1Default),
Array("no overlap",
- makeFeature("chrQ", 220, 230), pBam01, pFastq1, pFastq1Default),
+ makeInterval("chrQ", 220, 230), pBam01, pFastq1, pFastq1Default),
Array("partial overlap",
- makeFeature("chrQ", 430, 460), pBam01, pFastq1, pFastq1Default.updated("r04", true)),
+ makeInterval("chrQ", 430, 460), pBam01, pFastq1, pFastq1Default.updated("r04", true)),
Array("enveloped",
- makeFeature("chrQ", 693, 698), pBam01, pFastq1, pFastq1Default.updated("r03", true)),
+ makeInterval("chrQ", 693, 698), pBam01, pFastq1, pFastq1Default.updated("r03", true)),
Array("in intron",
- makeFeature("chrQ", 900, 999), pBam01, pFastq1, pFastq1Default.updated("r05", true))
+ makeInterval("chrQ", 900, 999), pBam01, pFastq1, pFastq1Default.updated("r05", true))
)
}
@Test(dataProvider = "pairAlnProvider1")
- def testPairBamDefault(name: String, feat: Feature, inAln: File,
+ def testPairBamDefault(name: String, feat: Interval, inAln: File,
fastqMap: Map[String, FastqPair], resultMap: Map[String, Boolean]) = {
require(resultMap.keySet == fastqMap.keySet)
val memFunc = makeMembershipFunction(Iterator(feat), inAln, commonSuffixLength = 2)
@@ -175,13 +186,13 @@ class ExtractAlignedFastqUnitTest extends TestNGSuite with MockitoSugar with Mat
}
@Test def testWriteSingleBamDefault() = {
- val memFunc = (recs: FastqPair) => Set("r01", "r03").contains(recs._1.getReadHeader)
- val in1 = new FastqReader(resourceFile("/single01.fq"))
- val mo1 = mock[BasicFastqWriter]
- selectFastqReads(memFunc, in1, mo1)
- verify(mo1, times(2)).write(anyObject.asInstanceOf[FastqRecord])
- verify(mo1).write(new FastqRecord("r01", "A", "", "H"))
- verify(mo1).write(new FastqRecord("r03", "G", "", "H"))
+ val memFunc = (recs: FastqPair) => Set("r01", "r03").contains(recs._1.getReadHeader)
+ val in1 = new FastqReader(resourceFile("/single01.fq"))
+ val mo1 = mock[BasicFastqWriter]
+ selectFastqReads(memFunc, in1, mo1)
+ verify(mo1, times(2)).write(anyObject.asInstanceOf[FastqRecord])
+ verify(mo1).write(new FastqRecord("r01", "A", "", "H"))
+ verify(mo1).write(new FastqRecord("r03", "G", "", "H"))
}
@Test def testWritePairBamDefault() = {