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package var2;
import java.io.PrintStream;
import java.util.ArrayList;
import java.util.HashMap;
import fileIO.ByteFile;
import fileIO.FileFormat;
import fileIO.ReadWrite;
import shared.Parse;
import shared.Parser;
import shared.PreParser;
import shared.Shared;
import shared.Timer;
import shared.Tools;
import stream.ConcurrentReadInputStream;
import stream.ConcurrentReadOutputStream;
import stream.FASTQ;
import stream.FastaReadInputStream;
import stream.Read;
import structures.ByteBuilder;
import structures.CoverageArray;
import structures.CoverageArray2;
import structures.ListNum;
import tracker.ReadStats;
/**
* Applies variants
*
* @author Brian Bushnell
* @date August 27, 2019
*
*/
public class ApplyVariants {
/*--------------------------------------------------------------*/
/*---------------- Initialization ----------------*/
/*--------------------------------------------------------------*/
/**
* Code entrance from the command line.
* @param args Command line arguments
*/
public static void main(String[] args){
//Start a timer immediately upon code entrance.
Timer t=new Timer();
//Create an instance of this class
ApplyVariants x=new ApplyVariants(args);
//Run the object
x.process(t);
//Close the print stream if it was redirected
Shared.closeStream(x.outstream);
}
/**
* Constructor.
* @param args Command line arguments
*/
public ApplyVariants(String[] args){
{//Preparse block for help, config files, and outstream
PreParser pp=new PreParser(args, getClass(), false);
args=pp.args;
outstream=pp.outstream;
}
//Set shared static variables prior to parsing
ReadWrite.USE_PIGZ=ReadWrite.USE_UNPIGZ=true;
ReadWrite.setZipThreads(Shared.threads());
Shared.capBuffers(4); //Only for singlethreaded programs
{//Parse the arguments
final Parser parser=parse(args);
Parser.processQuality();
maxReads=parser.maxReads;
overwrite=ReadStats.overwrite=parser.overwrite;
append=ReadStats.append=parser.append;
in1=parser.in1;
extin=parser.extin;
out1=parser.out1;
extout=parser.extout;
}
doPoundReplacement(); //Replace # with 1 and 2
adjustInterleaving(); //Make sure interleaving agrees with number of input and output files
fixExtensions(); //Add or remove .gz or .bz2 as needed
checkFileExistence(); //Ensure files can be read and written
checkStatics(); //Adjust file-related static fields as needed for this program
//Create output FileFormat objects
ffout1=FileFormat.testOutput(out1, FileFormat.FASTA, extout, true, overwrite, append, ordered);
//Create input FileFormat objects
ffin1=FileFormat.testInput(in1, FileFormat.FASTA, extin, true, true);
ffvcf=FileFormat.testInput(inVcf, FileFormat.VCF, null, true, true);
ffdepth=FileFormat.testInput(inDepth, FileFormat.TXT, null, true, false);
}
/*--------------------------------------------------------------*/
/*---------------- Initialization Helpers ----------------*/
/*--------------------------------------------------------------*/
/** Parse arguments from the command line */
private Parser parse(String[] args){
//Create a parser object
Parser parser=new Parser();
//Set any necessary Parser defaults here
//parser.foo=bar;
//Parse each argument
for(int i=0; i<args.length; i++){
String arg=args[i];
//Break arguments into their constituent parts, in the form of "a=b"
String[] split=arg.split("=");
String a=split[0].toLowerCase();
String b=split.length>1 ? split[1] : null;
if(b!=null && b.equalsIgnoreCase("null")){b=null;}
if(a.equals("verbose")){
verbose=Parse.parseBoolean(b);
}else if(a.equals("vcf") || a.equals("vars")){
inVcf=b;
}else if(a.equals("name") || a.equals("sample") || a.equals("samplename")){
sampleName=b;
}else if(a.equals("addnumbers")){
addContigNumbers=Parse.parseBoolean(b);
}else if(a.equals("useprefix") || a.equals("prefix")){
usePrefix=Parse.parseBoolean(b);
}else if(a.equals("delimiter")){
delimiter=Parse.parseSymbolToCharacter(b);
}else if(a.equals("cov") || a.equals("depth") || a.equals("indepth") || a.equals("basecov")){
inDepth=b;
}else if(a.equals("mindepth") || a.equals("mincov")){
minDepth=Integer.parseInt(b);
}else if(a.equals("maxindel")){
maxIndel=Integer.parseInt(b);
if(maxIndel<0){maxIndel=Integer.MAX_VALUE;}
}else if(a.equals("noindels")){
noIndels=Parse.parseBoolean(b);
}else if(a.equals("noframeshifts") || a.equals("banframeshifts")){
noFrameshifts=Parse.parseBoolean(b);
}else if(a.equals("frameshifts")){
noFrameshifts=!Parse.parseBoolean(b);
}else if(a.equals("parse_flag_goes_here")){
long fake_variable=Parse.parseKMG(b);
//Set a variable here
}else if(parser.parse(arg, a, b)){//Parse standard flags in the parser
//do nothing
}else{
outstream.println("Unknown parameter "+args[i]);
assert(false) : "Unknown parameter "+args[i];
}
}
return parser;
}
/** Replace # with 1 and 2 in headers */
private void doPoundReplacement(){
//Ensure there is an input file
if(in1==null || inVcf==null){throw new RuntimeException("Error - one sequence and one vcf file are required.");}
if(minDepth>0 && inDepth==null){throw new RuntimeException("Error - mindepth requires a coverage file.");}
}
/** Add or remove .gz or .bz2 as needed */
private void fixExtensions(){
in1=Tools.fixExtension(in1);
inVcf=Tools.fixExtension(inVcf);
inDepth=Tools.fixExtension(inDepth);
}
/** Ensure files can be read and written */
private void checkFileExistence(){
//Ensure output files can be written
if(!Tools.testOutputFiles(overwrite, append, false, out1)){
outstream.println((out1==null)+", "+out1);
throw new RuntimeException("\n\noverwrite="+overwrite+"; Can't write to output files "+out1+"\n");
}
//Ensure input files can be read
if(!Tools.testInputFiles(false, true, in1, inVcf, inDepth)){
throw new RuntimeException("\nCan't read some input files.\n");
}
//Ensure that no file was specified multiple times
if(!Tools.testForDuplicateFiles(true, in1, inVcf, out1)){
throw new RuntimeException("\nSome file names were specified multiple times.\n");
}
}
/** Make sure interleaving agrees with number of input and output files */
private void adjustInterleaving(){
FASTQ.FORCE_INTERLEAVED=false;
FASTQ.TEST_INTERLEAVED=false;
}
/** Adjust file-related static fields as needed for this program */
private static void checkStatics(){
//Adjust the number of threads for input file reading
if(!ByteFile.FORCE_MODE_BF1 && !ByteFile.FORCE_MODE_BF2 && Shared.threads()>2){
ByteFile.FORCE_MODE_BF2=true;
}
assert(FastaReadInputStream.settingsOK());
}
/*--------------------------------------------------------------*/
/*---------------- Outer Methods ----------------*/
/*--------------------------------------------------------------*/
/** Create read streams and process all data */
void process(Timer t){
VCFFile vfile=new VCFFile(ffvcf);
//Load vcf
if(ScafMap.defaultScafMap()==null){
ScafMap.setDefaultScafMap(vfile.toScafMap(null), ffvcf.name());
}
if(ffdepth!=null && minDepth>0){
depthMap=CoverageArray.loadDepth(ffdepth, CoverageArray2.class);
}
ArrayList<VCFLine> lines=vfile.lines(true);
varMap=new HashMap<String, ArrayList<Var>>(ScafMap.defaultScafMap().size());
for(VCFLine line : lines){
ArrayList<Var> value=varMap.get(line.scaf);
if(value==null){
value=new ArrayList<Var>();
varMap.put(line.scaf, value);
}
value.add(line.toVar());
}
//Create a read input stream
final ConcurrentReadInputStream cris=makeCris();
//Optionally create a read output stream
final ConcurrentReadOutputStream ros=makeCros();
//Reset counters
readsProcessed=readsOut=0;
basesProcessed=basesOut=0;
//Process the read stream
processInner(cris, ros);
if(verbose){outstream.println("Finished; closing streams.");}
//Write anything that was accumulated by ReadStats
errorState|=ReadStats.writeAll();
//Close the read streams
errorState|=ReadWrite.closeStreams(cris, ros);
//Report timing and results
t.stop();
outstream.println(Tools.timeReadsBasesProcessed(t, readsProcessed, basesProcessed, 8));
outstream.println(Tools.readsBasesOut(readsProcessed, basesProcessed, readsOut, basesOut, 8, false));
//Throw an exception of there was an error in a thread
if(errorState){
throw new RuntimeException(getClass().getName()+" terminated in an error state; the output may be corrupt.");
}
}
/*--------------------------------------------------------------*/
/*---------------- Inner Methods ----------------*/
/*--------------------------------------------------------------*/
private ConcurrentReadInputStream makeCris(){
ConcurrentReadInputStream cris=ConcurrentReadInputStream.getReadInputStream(maxReads, true, ffin1, null);
cris.start(); //Start the stream
if(verbose){outstream.println("Started cris");}
boolean paired=cris.paired();
if(!ffin1.samOrBam()){outstream.println("Input is being processed as "+(paired ? "paired" : "unpaired"));}
return cris;
}
private ConcurrentReadOutputStream makeCros(){
if(ffout1==null){return null;}
//Select output buffer size based on whether it needs to be ordered
final int buff=4;
final ConcurrentReadOutputStream ros=ConcurrentReadOutputStream.getStream(ffout1, null, buff, null, false);
ros.start(); //Start the stream
return ros;
}
/** Iterate through the reads */
void processInner(final ConcurrentReadInputStream cris, final ConcurrentReadOutputStream ros){
//Do anything necessary prior to processing
{
//Grab the first ListNum of reads
ListNum<Read> ln=cris.nextList();
//Check to ensure pairing is as expected
if(ln!=null && !ln.isEmpty()){
Read r=ln.get(0);
assert(ffin1.samOrBam() || (r.mate!=null)==cris.paired());
}
//As long as there is a nonempty read list...
while(ln!=null && ln.size()>0){
// if(verbose){outstream.println("Fetched "+reads.size()+" reads.");} //Disabled due to non-static access
processList(ln, cris, ros);
//Fetch a new list
ln=cris.nextList();
}
//Notify the input stream that the final list was used
if(ln!=null){
cris.returnList(ln.id, ln.list==null || ln.list.isEmpty());
}
}
//Do anything necessary after processing
}
/**
* Process a list of Reads.
* @param ln The list.
* @param cris Read Input Stream
* @param ros Read Output Stream for reads that will be retained
*/
void processList(ListNum<Read> ln, final ConcurrentReadInputStream cris, final ConcurrentReadOutputStream ros){
//Grab the actual read list from the ListNum
final ArrayList<Read> reads=ln.list;
//Loop through each read in the list
for(int idx=0; idx<reads.size(); idx++){
final Read r=reads.get(idx);
//Validate reads in worker threads
if(!r.validated()){r.validate(true);}
//Track the initial length for statistics
final int initialLength1=r.length();
//Increment counters
readsProcessed+=r.pairCount();
basesProcessed+=initialLength1;
Read mutant=processRead(r);
reads.set(idx, mutant);
readsOut+=mutant.pairCount();
basesOut+=mutant.pairLength();
}
//Output reads to the output stream
if(ros!=null){ros.add(reads, ln.id);}
//Notify the input stream that the list was used
cris.returnList(ln);
// if(verbose){outstream.println("Returned a list.");} //Disabled due to non-static access
}
@SuppressWarnings("unused")
private void applyDepth(Read r){applyDepth(r, null);}
private void applyDepth(Read r, CoverageArray ca){
if(ca==null){ca=depthMap.get(r.id);}
if(ca==null){
String s=Tools.trimToWhitespace(r.id);
ca=depthMap.get(s);
}
assert(ca!=null) : "Can't find "+r.id+" in depth map.";
final byte[] bases=r.bases;
for(int i=0; i<r.bases.length; i++){
if(ca.get(i)<minDepth){bases[i]=noCovSymbol;}
}
ArrayList<Var> vars=varMap.get(r.id);
if(vars==null){
String sub=Tools.trimToWhitespace(r.id);
vars=varMap.get(sub);//Handles truncated sequence names
}
if(vars==null){return;}
int removed=0;
for(int i=0; i<vars.size(); i++){
Var v=vars.get(i);
if(ca.get(v.start)<minDepth && v.indel()){
vars.set(i, null);
removed++;
}
}
if(removed>0){Tools.condenseStrict(vars);}
}
private void filterIndels(final Read r){
filterIndels(r, null);
}
private void filterIndels(final Read r, CoverageArray ca){
if(minDepth<=0 && maxIndel==Integer.MAX_VALUE && !noIndels && !noFrameshifts){return;}
if(minDepth>0 && ca==null){
ca=depthMap.get(r.id);
if(ca==null){
String s=Tools.trimToWhitespace(r.id);
ca=depthMap.get(s);
}
assert(ca!=null) : "Can't find "+r.id+" in depth map.";
}
ArrayList<Var> vars=varMap.get(r.id);
if(vars==null){
String sub=Tools.trimToWhitespace(r.id);
vars=varMap.get(sub);//Handles truncated sequence names
}
if(vars==null){return;}
int removed=0;
for(int i=0; i<vars.size(); i++){
Var v=vars.get(i);
boolean remove=false;
if(noFrameshifts && v.frameshift()){
remove=true;
}else if(v.indel()){
if(noIndels || Tools.max(v.reflen(), v.readlen())>maxIndel
|| (ca!=null && ca.get(v.start)<minDepth))
remove=true;
}
if(remove){
vars.set(i, null);
removed++;
}
}
if(removed>0){Tools.condenseStrict(vars);}
}
/**
* Process a single read.
* @param r Read 1
*/
Read processRead(final Read r){
CoverageArray ca=null;
if(minDepth>0){
assert(depthMap!=null) : "minDepth is "+minDepth+" but depthMap is null. "
+ "You need a coverage file which includes this contig.";
ca=depthMap.get(r.id);
if(ca==null){
String s=Tools.trimToWhitespace(r.id);
ca=depthMap.get(s);
}
assert(ca!=null) : "Can't find "+r.id+" in depth map.";
applyDepth(r, ca);
}
filterIndels(r, ca);
String name=r.id;
final byte[] bases=r.bases;
ArrayList<Var> vars=varMap.get(name);
if(vars==null){return r;}
HashMap<Integer, Var> map=new HashMap<Integer, Var>();
for(Var v : vars){
Integer key=v.start;
Var old=map.get(key);
if(old==null || old.alleleCount()<v.alleleCount()){//TODO: allow retaining the higher quality one
map.put(key, v);
}
}
ByteBuilder bb=new ByteBuilder();
for(int i=0; i<bases.length; ){
Var v=map.get(i);
if(v==null){
bb.append(bases[i]);
i++;
}else{
final int reflen=v.reflen();
final int readlen=v.readlen();
applied++;
//Old code, did not handle 'N' for low coverage
// if(readlen>0){
// bb.append(v.allele);
// }
// if(reflen==0){//insertion
// bb.append(bases[i]);
// i++;
// }else{
// i+=reflen;
// }
//low-depth indels should have already been removed
if(reflen==0){//insertion
// final int depth2=(ca==null || minDepth<1 ? 9999 : ca.get(i));
// if(depth2>=minDepth){//Apply ins
// bb.append(v.allele);
// }
bb.append(bases[i]);
i++;
}else if(readlen==0){//del
// final int depth2=(ca==null || minDepth<1 ? 9999 : ca.get(i));
// if(depth2>=minDepth){//Apply del
i+=reflen;
// }else{//Ignore del
// bb.append(bases[i]);
// i++;
// }
}else if(readlen!=reflen){//complex
int lim=i+reflen;
for(int j=0; i<lim; i++, j++){
final int depth2=(ca==null || minDepth<1 ? 9999 : ca.get(i));
byte b=(j<v.allele.length ? v.allele[j] : bases[i]);
bb.append(depth2>=minDepth ? b : (byte)'N');
}
}else{//sub
assert(v.reflen()==v.readlen()) : v;
for(byte b : v.allele){
final int depth2=(ca==null || minDepth<1 ? 9999 : ca.get(i));
bb.append(depth2>=minDepth ? b : (byte)'N');
i++;
}
}
}
}
final String name2=rename(name, r.numericID);
Read mutant=new Read(bb.toBytes(), null, name2, r.numericID);
return mutant;
}
private String rename(String old, long number){
if(sampleName==null){return old;}
nameBuilder.clear();
if(usePrefix){
return nameBuilder.append(sampleName).append(delimiter).append(old).toString();
}else if(addContigNumbers){
return nameBuilder.append(sampleName).append(delimiter).append(number).toString();
}
return sampleName;
}
/*--------------------------------------------------------------*/
/*---------------- Fields ----------------*/
/*--------------------------------------------------------------*/
/** Primary input file path */
private String in1=null;
/** Variant input file */
private String inVcf;
/** Per-base coverage depth file (optional) */
private String inDepth;
/** Primary output file path */
private String out1=null;
/** Override input file extension */
private String extin=null;
/** Override output file extension */
private String extout=null;
/** If positive, change regions below this depth to N */
private int minDepth=0;
private byte noCovSymbol='N';
HashMap<String, ArrayList<Var>> varMap;
HashMap<String, CoverageArray> depthMap;
/** Name of output sequences, if different from input */
String sampleName=null;
/** Add numbers as a suffix to the name of each contig, if they will be renamed */
boolean addContigNumbers=true;
/** Use samplename as prefix, rather than replacing the existing name. */
boolean usePrefix=false;
/** Delimits modified names. */
char delimiter='_';
/** Ignore non-multiple-of-3 indels */
private boolean noFrameshifts=false;
/** Ignore indels longer than this */
private int maxIndel=Integer.MAX_VALUE;
/** Ignore indels */
private boolean noIndels=false;
private ByteBuilder nameBuilder=new ByteBuilder();
/*--------------------------------------------------------------*/
/** Number of reads processed */
protected long readsProcessed=0;
/** Number of bases processed */
protected long basesProcessed=0;
/** Number of reads retained */
protected long readsOut=0;
/** Number of bases retained */
protected long basesOut=0;
/** Quit after processing this many input reads; -1 means no limit */
private long maxReads=-1;
private long applied=0;
/*--------------------------------------------------------------*/
/*---------------- Final Fields ----------------*/
/*--------------------------------------------------------------*/
/** Primary input file */
private final FileFormat ffin1;
/** Variant input file */
private final FileFormat ffvcf;
/** Coverage input file */
private final FileFormat ffdepth;
/** Primary output file */
private final FileFormat ffout1;
/*--------------------------------------------------------------*/
/*---------------- Common Fields ----------------*/
/*--------------------------------------------------------------*/
/** Print status messages to this output stream */
private PrintStream outstream=System.err;
/** Print verbose messages */
public static boolean verbose=false;
/** True if an error was encountered */
public boolean errorState=false;
/** Overwrite existing output files */
private boolean overwrite=true;
/** Append to existing output files */
private boolean append=false;
/** This flag has no effect on singlethreaded programs */
private final boolean ordered=false;
}
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