workflow pancancer_controlsample{
	String normal
	String tumor
	String cancer
	String project = "650gene"
	String bed = "/dataseq/jmdna/database/bed/650.bed"
	String bed_plus = "/dataseq/jmdna/database/bed/650plus50bp.bed"
	String name
	String outputDir
	String need_TMB_test = "yes"
	String need_MSI_test = "yes"
	String need_neoantigen_test = "yes"
	String inputDir	
	String ref = "/dataseq/jmdna/database/genome/hg19/hg19.fa"
	String codes_dir = "/dataseq/jmdna/codes/pancancer_controlsample"
	String accessBed = "/dataseq/jmdna/software/cnvkit-0.9.7/data/access-5k-mappable.hg19.bed"
	String annotateGene = "/dataseq/jmdna/software/cnvkit-0.9.7/data/refFlat.txt"
	String gc_wiggle = "/dataseq/jmdna/codes/pancancer_controlsample/hg19.gc200Base.txt.gz"
#创建目录
    call create_dir{
        input:
            outputDir=outputDir,
	    need_neoantigen_test=need_neoantigen_test,
	    need_MSI_test=need_MSI_test
    }
	scatter (name in [normal,tumor]){
	call qc{
        input:
            name=name,
            inputDir=inputDir,
            outputDir=outputDir
    }
    call alignment_bwa{
        input:
            name=name,
            outputDir=outputDir,
            ref=ref,
            read1=qc.outputFile[0],
            read2=qc.outputFile[1]
    }
    call markduplicates{
        input:
            name=name,
            outputDir=outputDir,
            ref=ref,
            sortedBam=alignment_bwa.sorted
    }	
	call generater_mpileup{
		input:
			name=name,
			ref=ref,
			bed=bed_plus,
			outputDir=outputDir,
			rmdupBam=markduplicates.rmdup
	}

	}
	
	call mutation_calling{
        input:
		codes_dir=codes_dir,
		tumor=tumor,
 		outputDir=outputDir,			
 		pileup=generater_mpileup.pileup
    }
	
    call annovar{
 	input:
		tumor=tumor,
		normal=normal,
		outputDir=outputDir,
		ref=ref,
		vcf=mutation_calling.vcf
    }
	scatter (name in [normal,tumor]){	
		call qc_2{
			input:
				ref=ref,
				bed=bed,
				codes_dir=codes_dir,
				name=name,
				tumor=tumor,
				normal=normal,
				outputDir=outputDir,
				anno=annovar.anno
		}
	}	
    call dealwithsnvindel{
	input:
		codes_dir=codes_dir,
		project=project,
		outputDir=outputDir,
		tumor=tumor,
		cancer=cancer,
		anno=annovar.anno
    }
	call fusion{
		input:
			ref=ref,
			codes_dir=codes_dir,
                        project=project,
			tumor=tumor,
			outputDir=outputDir,
			cancer=cancer,
			rmdupBam=markduplicates.rmdup
	}
	
	if (need_TMB_test=="yes"){
	call TMB{
		input:
			codes_dir=codes_dir,
			tumor=tumor,
			outputDir=outputDir,
			anno=annovar.anno,
			concordance=conpair.concordance,
			snv_result=dealwithsnvindel.snv,
			cnv_result=cnvkit.cnv,
			fusion_result=fusion.fusion
	}
	}
	
	if (need_MSI_test=="yes"){
	call MSI{
		input:
			bed=bed,
			tumor=tumor,
			normal=normal,
			outputDir=outputDir,
			rmdupBam=markduplicates.rmdup
	}
	}
    call MMR{
		input:
			codes_dir=codes_dir,
			tumor=tumor,
			outputDir=outputDir,
			snv=dealwithsnvindel.snv
	}
	call HRR{
		input:
			codes_dir=codes_dir,
			tumor=tumor,
			outputDir=outputDir,
			snv=dealwithsnvindel.snv
	}
	call chemoTherapy{
		input:
			codes_dir=codes_dir,
			normal=normal,
			tumor=tumor,
			outputDir=outputDir,
			ref=ref,
			project=project,
			rmdupBam=markduplicates.rmdup
	}
	
	call hereditary{
		input:		
			codes_dir=codes_dir,
			tumor=tumor,
			outputDir=outputDir,
			project=project,
			snv=dealwithsnvindel.snv
	}

	call conpair{
		input:
			name=name,
			codes_dir=codes_dir,
			tumor=tumor,
			normal=normal,
			ref=ref,
			outputDir=outputDir,
			rmdupBam=markduplicates.rmdup
	}
	call cnvkit{
        input:
		name=name,
            	normal=normal,
            	tumor=tumor,
            	ref=ref,
            	bed=bed,
            	cancer=cancer,
           	project=project,
            	codes_dir=codes_dir,
           	outputDir=outputDir,
            	rmdupBam=markduplicates.rmdup,
            	accessBed=accessBed,
            	annotateGene=annotateGene,
		purity=tumor_content.purity
    }
	call tumor_content{
		input:
			name=name,
			ref=ref,
			tumor=tumor,
			normal=normal,
			outputDir=outputDir,
			gc_wiggle=gc_wiggle,
			codes_dir=codes_dir,
			pileup=generater_mpileup.pileup
	}

	if (need_neoantigen_test=="yes"){
	call HLA{
		input:
			inputDir=inputDir,
			outputDir=outputDir,
			normal=normal,
			newdir=create_dir.newdir
	}
	call neoantigen{
		input:
			codes_dir=codes_dir,
			outputDir=outputDir,
			name=name,
			normal=normal,
			tumor=tumor,
			hla=HLA.hla,
			vcf=mutation_calling.vcf
	}
	}
	
	call auto_report{
		input:
			cancer=cancer,
			codes_dir=codes_dir,
			outputDir=outputDir,
			name=name,
			normal=normal,
			tumor=tumor,	
			qc_2=qc_2.qc,
			concordance=conpair.concordance,
			snv_result=dealwithsnvindel.snv,
			cnv_result=cnvkit.cnv,
			fusion_result=fusion.fusion,
			tmb=TMB.tmb
	}
	call hotspot{
		input:
			tumor=tumor,
			outputDir=outputDir,
			ref=ref,
			rmdupBam=markduplicates.rmdup,
			codes_dir=codes_dir
        }

}
#create project directory
task create_dir{
    String outputDir
	String need_MSI_test
	String need_neoantigen_test
    command <<<
        #创建目录
        if [ ! -d ${outputDir} ];then
            mkdir ${outputDir}
        fi

        #创建qc目录
        if [ ! -d ${outputDir}/qc ];then
            mkdir ${outputDir}/qc
        fi

        #创建alignment目录
        if [ ! -d ${outputDir}/alignment ];then
            mkdir ${outputDir}/alignment
        fi

        #创建mutation目录
        if [ ! -d ${outputDir}/mutation ];then
            mkdir ${outputDir}/mutation
        fi

        #创建cnv目录
        if [ ! -d ${outputDir}/cnvkit ];then
           mkdir ${outputDir}/cnvkit
        fi

		#创建report目录
        if [ ! -d ${outputDir}/report ];then
            mkdir -p ${outputDir}/report
        fi
		
		#创建chemo目录
        if [ ! -d ${outputDir}/chemo ];then
            mkdir ${outputDir}/chemo
        fi

        #创建fusion目录
        if [ ! -d ${outputDir}/fusion ];then
            mkdir ${outputDir}/fusion
        fi

	#创建MSI目录
	if [ ${need_MSI_test} = "yes" ] && [ ! -d ${outputDir}/MSI ];then
		mkdir ${outputDir}/MSI
	fi
		
		
	if [ ${need_neoantigen_test} = "yes" ] && [ ! -d ${outputDir}/neoantigen/HLA ];then
		mkdir -p ${outputDir}/neoantigen/HLA
	fi
		
	#创建MMR目录
	if [ ! -d ${outputDir}/MMR ];then
		mkdir ${outputDir}/MMR
	fi

	##创建HRR目录
	if [ ! -d ${outputDir}/HRR ];then
        	mkdir ${outputDir}/HRR
        fi

        #创建HPD目录
        if [ ! -d ${outputDir}/HPD ];then
            mkdir ${outputDir}/HPD
        fi

    >>>
	output{
          String newdir = "${outputDir}/report"
      }
}

#generator raw fastq to clean fastq
task qc{
    String name
    String inputDir
    String outputDir

    command <<<

        echo processing raw reads with fastp
        fastp -i ${inputDir}/*_${name}_*1.fq.gz -o ${outputDir}/qc/${name}_clean_R1.fq.gz \
        -I ${inputDir}/*_${name}_*2.fq.gz -O ${outputDir}/qc/${name}_clean_R2.fq.gz \
        -w 10 \
	--correction \
        --overlap_len_require 10 \
        -j ${outputDir}/qc/${name}.json \
        -h ${outputDir}/qc/${name}.html --report_title ${name} -e 20

#	--adapter_sequence AAGTCGGAGGCCAAGCGGTCTTAGGAAGACAA \
#	--adapter_sequence_r2 AAGTCGGATCGTAGCCATGTCGTTCTGTGAGCCAAGGAGTTG \


    >>>

    output{
        Array[String] outputFile = [
        "${outputDir}/qc/${name}_clean_R1.fq.gz ",
        "${outputDir}/qc/${name}_clean_R2.fq.gz",
        "${outputDir}/qc/${name}.json",
        "${outputDir}/qc/${name}.html"
        ]
    }
}

#alignment clean fastq to reference
task alignment_bwa{
      String name
      String ref
      String outputDir
      String read1
      String read2
      command<<<
 
         bwa mem -R '@RG\tID:group_n\tLB:library_n\tPL:BGI\tPU:unit1\tSM:${name}' -M -t 10 ${ref} \
          ${outputDir}/qc/${name}_clean_R1.fq.gz ${outputDir}/qc/${name}_clean_R2.fq.gz | \
          samtools view -@ 5 -bh  -o - | samtools sort -@ 5  -o ${outputDir}/alignment/${name}.sorted.bam

          samtools index ${outputDir}/alignment/${name}.sorted.bam

      >>>

      output{
          String sorted = "${outputDir}/alignment/${name}.sorted.bam"
      }
}

#remove PCR duplicates 
task markduplicates{
    String name
    String ref
    String outputDir
    String sortedBam

    command<<<
 
       java -XX:+UseParallelGC -XX:ParallelGCThreads=2 -Xmx12G  -jar $PICARD MarkDuplicates \
        I=${outputDir}/alignment/${name}.sorted.bam \
        O=${outputDir}/alignment/${name}.rmdup.bam  \
        CREATE_INDEX=true \
        M=${outputDir}/alignment/${name}.rmdup.metrics.txt  \
        R=${ref}

    >>>

    output{
              String  rmdup = "${outputDir}/alignment/${name}.rmdup.bam"
    }
}

# generater mpileup file
task generater_mpileup{
    String name
    String ref
    String bed
    String outputDir
	String rmdupBam

    command<<<

        samtools mpileup -Bq 20 -Q 20  -f ${ref} -l ${bed} \
        ${outputDir}/alignment/${name}.rmdup.bam -o ${outputDir}/alignment/${name}.pileup

    >>>

    output{
        String pileup = "${outputDir}/alignment/${name}.pileup"
    }
}

task mutation_calling{
	String codes_dir
	String tumor
	Array[String] pileup
	String outputDir
	
	command<<<
	
		sh ${codes_dir}/mpileup2cns.sh ${sep=" " pileup} ${outputDir} ${tumor}
		java -jar $GATK MergeVcfs -I ${outputDir}/mutation/${tumor}.snp.Somatic.hc.vcf -I ${outputDir}/mutation/${tumor}.indel.Somatic.hc.vcf \
		-O ${outputDir}/mutation/${tumor}.snp.indel.Somatic.hc.vcf -D /dataseq/jmdna/database/genome/hg19/hg19.dict
		java -jar $GATK MergeVcfs -I ${outputDir}/mutation/${tumor}.snp.Germline.vcf -I ${outputDir}/mutation/${tumor}.indel.Germline.vcf \
		-O ${outputDir}/mutation/${tumor}.snp.indel.Germline.vcf -D /dataseq/jmdna/database/genome/hg19/hg19.dict
		java -jar $GATK MergeVcfs -I ${outputDir}/mutation/${tumor}.snp.LOH.hc.vcf -I ${outputDir}/mutation/${tumor}.indel.LOH.hc.vcf \
		-O ${outputDir}/mutation/${tumor}.snp.indel.LOH.hc.vcf -D /dataseq/jmdna/database/genome/hg19/hg19.dict

	>>>
	
	output{
        Array[String] vcf = [
        "${outputDir}/mutation/${tumor}.snp.indel.Somatic.hc.vcf",
        "${outputDir}/mutation/${tumor}.snp.indel.Germline.vcf",
        "${outputDir}/mutation/${tumor}.snp.indel.LOH.hc.vcf"
        ]
    }
}


task annovar{
	String tumor
	String normal
    	String outputDir
	String ref
   	Array[String] vcf
    command<<<

        table_annovar.pl \
        ${outputDir}/mutation/${tumor}.snp.indel.Somatic.hc.vcf \
        /dataseq/jmdna/software/annovar/humandb/ \
        -buildver hg19 -nastring . -vcfinput -remove -otherinfo \
        -protocol refGene,avsnp150,cosmic91,clinvar_20220320,1000g2015aug_all,1000g2015aug_eas,esp6500siv2_all,exac03nontcga,gnomad_genome,dbnsfp35c,cytoBand \
		-argument '-splicing_threshold 2 -hgvs',,,,,,,,,, \
        --intronhgvs 50 \
        -operation g,f,f,f,f,f,f,f,f,f,r \
		--outfile ${outputDir}/mutation/${tumor}.snp.indel.Somatic.anno

        table_annovar.pl \
        ${outputDir}/mutation/${tumor}.snp.indel.Germline.vcf \
        /dataseq/jmdna/software/annovar/humandb/ \
        -buildver hg19 -nastring . -vcfinput -remove -otherinfo \
		-protocol refGene,avsnp150,cosmic91,clinvar_20220320,1000g2015aug_all,1000g2015aug_eas,esp6500siv2_all,exac03nontcga,gnomad_genome,dbnsfp35c,cytoBand \
        -argument '-splicing_threshold 2 -hgvs',,,,,,,,,, \
        --intronhgvs 50 \
		-operation g,f,f,f,f,f,f,f,f,f,r \
		--outfile ${outputDir}/mutation/${tumor}.snp.indel.Germline.anno

        table_annovar.pl \
        ${outputDir}/mutation/${tumor}.snp.indel.LOH.hc.vcf \
        /dataseq/jmdna/software/annovar/humandb/ \
        -buildver hg19 -nastring . -vcfinput -remove -otherinfo \
		-protocol refGene,avsnp150,cosmic91,clinvar_20220320,1000g2015aug_all,1000g2015aug_eas,esp6500siv2_all,exac03nontcga,gnomad_genome,dbnsfp35c,cytoBand \
        -argument '-splicing_threshold 2 -hgvs',,,,,,,,,, \
        --intronhgvs 50 \
		-operation g,f,f,f,f,f,f,f,f,f,r \
		--outfile ${outputDir}/mutation/${tumor}.snp.indel.LOH.anno

		java -jar /dataseq/jmdna/software/GenomeAnalysisTK.3.7.jar -T VariantAnnotator \
		-R ${ref} \
		-I ${outputDir}/alignment/${tumor}.rmdup.bam \
		-V ${outputDir}/mutation/${tumor}.snp.indel.Somatic.hc.vcf \
		-o ${outputDir}/mutation/${tumor}.TandemRepeatAnnotator.vcf \
		--annotation TandemRepeatAnnotator

# -nt 10

	grep -v "^##" ${outputDir}/mutation/${tumor}.TandemRepeatAnnotator.vcf | cut -f8| paste ${outputDir}/mutation/${tumor}.snp.indel.Somatic.anno.hg19_multianno.txt  -  >${outputDir}/mutation/${tumor}.snp.indel.Somatic.annoall.hg19_multianno.txt

    >>>
	output{
        Array[String] anno = [
        "${outputDir}/mutation/${tumor}.snp.indel.Somatic.anno.hg19_multianno.txt",
        "${outputDir}/mutation/${tumor}.snp.indel.Germline.anno.hg19_multianno.txt",
        "${outputDir}/mutation/${tumor}.snp.indel.Somatic.annoall.hg19_multianno.txt",
        ]
    }
}

task dealwithsnvindel{
	String codes_dir
	String project
	String outputDir
	String tumor
	Array[String] anno
	String cancer

	command <<<
	
	perl ${codes_dir}/pick_variant.pl ${outputDir} ${tumor}
	perl ${codes_dir}/pick_mut_splice_promoter.pl ${codes_dir} ${tumor} ${outputDir} ${project}
	perl /home/jm001/test_duantao/database_update/codes/682/targetTherapy.pl ${tumor} ${outputDir} ${project} ${cancer}
	perl /home/jm001/test_duantao/database_update/codes/682/germline_targetTherapy.pl ${tumor} ${outputDir} ${project} ${cancer}
	>>>
    output{
        String snv = "${outputDir}/mutation/${tumor}.snvindel.pos.txt"
    }
}


task qc_2{
	String ref
	String codes_dir
	String name
	String tumor
	String normal
	String bed
	String outputDir
	#for task chain
	Array[String] anno
	
	command <<<

		samtools flagstat -@ 10 ${outputDir}/alignment/${name}.rmdup.bam >${outputDir}/qc/${name}.rmdup.flagstat
		samtools stats --reference ${ref} -t ${bed} -@ 10 ${outputDir}/alignment/${name}.rmdup.bam > ${outputDir}/alignment/${name}.stat
		mkdir ${outputDir}/qc/${name}_bamdst
		bamdst -p ${bed} -o ${outputDir}/qc/${name}_bamdst ${outputDir}/alignment/${name}.rmdup.bam
		Rscript ${codes_dir}/InsertAndDepthStat.R ${outputDir}/qc/${name}_InsertAndDepthStat ${outputDir}/qc/${name}_bamdst/insertsize.plot ${outputDir}/qc/${name}_bamdst/depth_distribution.plot
		python3 ${codes_dir}/sample_post.py -s ${tumor} -o ${outputDir} --n ${normal}
		python3 ${codes_dir}/qc_stat.py ${outputDir}/qc/${name}.json ${outputDir}/qc/${name}_bamdst/coverage.report ${outputDir}/qc/${name}_bamdst/depth_distribution.plot ${outputDir} ${name} ${tumor}

	>>>
	output{
		String qc = "${outputDir}/qc/${name}_qc.txt"
	}
}

task fusion{
	String ref
	String codes_dir
	String tumor
	String outputDir
	String cancer
	String project
	#for task chain
	Array[String] rmdupBam

	command<<<
		# Extract the discordant paired-end alignments.
		samtools view -b -F 1294 ${outputDir}/alignment/${tumor}.rmdup.bam > ${outputDir}/fusion/${tumor}.discordants.bam

		# Extract the split-read alignments
		samtools view -h ${outputDir}/alignment/${tumor}.rmdup.bam \
		| /dataseq/jmdna/software/lumpy-sv/scripts/extractSplitReads_BwaMem -i stdin \
		| samtools view -Sb - \
		> ${outputDir}/fusion/${tumor}.splitters.bam

		lumpyexpress \
		-B ${outputDir}/alignment/${tumor}.rmdup.bam \
		-S ${outputDir}/fusion/${tumor}.splitters.bam \
		-D ${outputDir}/fusion/${tumor}.discordants.bam \
		-o ${outputDir}/fusion/${tumor}.fusion.vcf

		perl ${codes_dir}/fusion.filter.pl ${outputDir}/fusion/${tumor}.fusion.vcf ${outputDir}/fusion/${tumor}.fusion.filter.vcf

		svtyper \
		-B ${outputDir}/alignment/${tumor}.rmdup.bam \
		-i ${outputDir}/fusion/${tumor}.fusion.filter.vcf \
		-T ${ref} \
		-o ${outputDir}/fusion/${tumor}.fusion.gt.vcf

		table_annovar.pl \
		${outputDir}/fusion/${tumor}.fusion.gt.vcf \
		/dataseq/jmdna/software/annovar/humandb/ \
		-buildver hg19 -nastring . -vcfinput -remove -otherinfo \
		-protocol refGene \
		-operation g \
		--outfile ${outputDir}/fusion/${tumor}.fusion
	
		perl ${codes_dir}/fusion.reanno.pl ${outputDir}/qc/${tumor}_bamdst/depth.tsv.gz ${outputDir} ${tumor}
		perl /home/jm001/test_duantao/database_update/codes/682/fusion_targetTherapy.pl ${codes_dir} ${tumor} ${outputDir} ${project} ${cancer}

	>>>

	output{
        	String fusion = "${outputDir}/fusion/${tumor}.fusion.pos.txt"
    	}
}

task chemoTherapy{
		String codes_dir
		String normal
		String tumor
		String outputDir
		String ref
		String project
		Array[String] rmdupBam
		command <<<
			# perl ${codes_dir}/chemo/singlecancer_chemo_2.pl ${codes_dir} ${outputDir} ${normal} ${ref} ${project}
			${codes_dir}/chemo/chemo_panel.py -p ${project} -o ${outputDir} --n ${normal}
        >>>
}

task hereditary{
		String codes_dir
		String tumor
		String outputDir
		String project
		String snv
		command <<<
			${codes_dir}/hereditary/hereditary.py -p ${project} -o ${outputDir} --n ${tumor}
        >>>
}



task conpair{
	String name
	String tumor
	String normal
	String outputDir
	String codes_dir
	String ref
	#for task chain
	Array[String] rmdupBam
	command <<<

		python3 /dataseq/jmdna/software/Conpair-master/scripts/run_gatk_pileup_for_sample.py \
		-M /dataseq/jmdna/software/Conpair-master/data/markers/GRCh37.autosomes.phase3_shapeit2_mvncall_integrated.20130502.SNV.genotype.sselect_v4_MAF_0.4_LD_0.8.bed \
		-B ${outputDir}/alignment/${normal}.rmdup.bam \
		-O ${outputDir}/alignment/${normal}.gatk.mpileup \
		-R ${ref} \
		-G /dataseq/jmdna/software/GenomeAnalysisTK.3.7.jar
		
		python3 /dataseq/jmdna/software/Conpair-master/scripts/run_gatk_pileup_for_sample.py \
		-M /dataseq/jmdna/software/Conpair-master/data/markers/GRCh37.autosomes.phase3_shapeit2_mvncall_integrated.20130502.SNV.genotype.sselect_v4_MAF_0.4_LD_0.8.bed \
		-B ${outputDir}/alignment/${tumor}.rmdup.bam \
		-O ${outputDir}/alignment/${tumor}.gatk.mpileup \
		-R ${ref} \
		-G /dataseq/jmdna/software/GenomeAnalysisTK.3.7.jar
	
		sed -i 's/^chr//g' ${outputDir}/alignment/${normal}.gatk.mpileup
		sed -i 's/^chr//g' ${outputDir}/alignment/${tumor}.gatk.mpileup

			
		python3 /dataseq/jmdna/software/Conpair-master/scripts/verify_concordance.py \
		-T ${outputDir}/alignment/${tumor}.gatk.mpileup \
		-N ${outputDir}/alignment/${normal}.gatk.mpileup \
		-H \
		-O ${outputDir}/qc/${name}_concordance.txt
		
		python3 /dataseq/jmdna/software/Conpair-master/scripts/estimate_tumor_normal_contamination.py \
		-T ${outputDir}/alignment/${tumor}.gatk.mpileup \
		-N ${outputDir}/alignment/${normal}.gatk.mpileup \
		-O ${outputDir}/qc/${name}_contamination.txt				

	>>>
	output{
		Array[String] concordance = [
        "${outputDir}/report/qc/${name}_concordance.txt",
		"${outputDir}/report/qc/${name}_contamination.txt",
		]
	}
}

task MSI{
		String bed
		String tumor
		String normal
		String outputDir
		Array[String] rmdupBam

		command <<<

			msisensor2 msi -d /dataseq/jmdna/software/msisensor2/hg19.microsatellites.list -n ${outputDir}/alignment/${normal}.rmdup.bam -t ${outputDir}/alignment/${tumor}.rmdup.bam -e ${bed} -b 10 -o ${outputDir}/MSI/${tumor}.msi

		>>>
		output{
			String target="${outputDir}/MSI/${tumor}.msi"
		}
}

task MMR{
	String codes_dir
	String tumor
	String outputDir
	String snv
	command <<<
			perl ${codes_dir}/mmr_controlsample.pl ${outputDir} ${tumor}
	>>>
	output{
			String mmr_out = "${outputDir}/MMR/${tumor}_mmr.txt"
		}
}

task HRR{
	String codes_dir
	String tumor
	String outputDir
	String snv
	command <<<
			perl ${codes_dir}/hrr_controlsample_tissue.pl ${outputDir} ${tumor}
	>>>
	output{
			String mmr_out = "${outputDir}/HRR/${tumor}_hrr.txt"
		}
}

task TMB{
	String codes_dir
	String tumor
	String outputDir
	Array[String] anno
	Array[String] concordance
	String snv_result
	String cnv_result
	String fusion_result



	
	
	command <<<	
		perl ${codes_dir}/tmb.pl ${outputDir} ${tumor}
	>>>
	output{
		String tmb="${outputDir}/mutation/${tumor}.tmb.txt"
	}
}

task cnvkit{
    String ref
    String bed
	String name
    String tumor
    String normal
    String outputDir
    String cancer
    String accessBed
    String annotateGene	
    String codes_dir
    String project
    #for task chain
    Array[String] rmdupBam
	String purity

    command <<<

	echo run cnvkit batch to processing cnv calling
        cnvkit.py batch \
        ${outputDir}/alignment/${tumor}.rmdup.bam \
        --normal ${outputDir}/alignment/${normal}.rmdup.bam \
        --targets ${bed} \
        --fasta ${ref} \
        --access ${accessBed} \
        --output-reference ${outputDir}/cnvkit/${normal}_reference.cnn \
        --annotate ${annotateGene} \
        --drop-low-coverage --scatter --output-dir ${outputDir}/cnvkit

	cnvkit.py scatter \
	${outputDir}/cnvkit/${tumor}.rmdup.cnr -s ${outputDir}/cnvkit/${tumor}.rmdup.cns \
	--y-max 3 --y-min -3 \
	--title ${tumor}.cns \
	-o ${outputDir}/cnvkit/${tumor}.cnv.png

    if [ -e "${outputDir}/qc/sequenza/${name}_confints_CP.txt" ]; then
        # absolute copy number
        cnvkit.py call \
        -m clonal \
        ${outputDir}/cnvkit/${tumor}.rmdup.cns \
        -y \
        --purity `head -n2 ${outputDir}/qc/sequenza/${name}_confints_CP.txt |tail -n1|cut -f1` \
        --drop-low-coverage  \
        --filter ampdel \
        -o ${outputDir}/cnvkit/${tumor}.rmdup.cns.cn.hc
    fi        
	perl ${codes_dir}/log2_cn.pl ${outputDir}/cnvkit/${tumor}.rmdup.cns ${outputDir}/cnvkit/${tumor}.rmdup.cns.cn	
	perl /home/jm001/test_duantao/database_update/codes/682/cnv_targetTherapy.pl ${codes_dir} ${tumor} ${outputDir} ${project} ${cancer}
    >>>

	output{
        String cnv = "${outputDir}/cnvkit/${tumor}.rmdup.cns"
    	}

}

task tumor_content{
	String name
	String ref
	String tumor
	String normal
	String outputDir
	String gc_wiggle
	String codes_dir
	#for task chain
	Array[String] pileup
	
	command <<<

		sequenza-utils bam2seqz -p -gc ${gc_wiggle} \
		-F ${ref} \
		-n ${outputDir}/alignment/${normal}.pileup \
		-t ${outputDir}/alignment/${tumor}.pileup | gzip >${outputDir}/qc/target_${name}.200base.seqz.gz
		sequenza-utils seqz_binning -w 200 -s ${outputDir}/qc/target_${name}.200base.seqz.gz | gzip  > ${outputDir}/qc/target_${name}.200base.small.seqz.gz

        Rscript ${codes_dir}/sequenza.R ${name} ${outputDir}/qc/target_${name}.200base.small.seqz.gz ${outputDir}/qc/sequenza || echo "sequenza failed!"
		
		
#		Rscript ${codes_dir}/qc_stat.R ${outputDir} ${name}
#		cp ${outputDir}/qc/${name}_qcstat.txt ${outputDir}/report/qc/${name}_qcstat.txt

	>>>
	output{
        String purity = "${outputDir}/qc/sequenza/${name}_CP_contours.pdf"
    }

}

task HLA{
	String inputDir
	String outputDir
	String normal
	String newdir
	command <<<

		razers3 -tc 10 -i 95 -m 1 -dr 0 -o ${outputDir}/neoantigen/HLA/fished_1.bam /dataseq/jmdna/software/OptiType-1.3.5/data/hla_reference_dna.fasta ${inputDir}/*_${normal}_*1.fq.gz
		samtools bam2fq ${outputDir}/neoantigen/HLA/fished_1.bam > ${outputDir}/neoantigen/HLA/${normal}_1_fished.fastq
		rm ${outputDir}/neoantigen/HLA/fished_1.bam 
		razers3 -tc 10 -i 95 -m 1 -dr 0 -o ${outputDir}/neoantigen/HLA/fished_2.bam /dataseq/jmdna/software/OptiType-1.3.5/data/hla_reference_dna.fasta ${inputDir}/*_${normal}_*2.fq.gz
		samtools bam2fq ${outputDir}/neoantigen/HLA/fished_2.bam > ${outputDir}/neoantigen/HLA/${normal}_2_fished.fastq
		rm ${outputDir}/neoantigen/HLA/fished_2.bam
		/dataseq/jmdna/software/OptiType-1.3.5/OptiTypePipeline.py -i ${outputDir}/neoantigen/HLA/${normal}_1_fished.fastq  ${outputDir}/neoantigen/HLA/${normal}_2_fished.fastq --dna -v --prefix ${normal} -o ${outputDir}/neoantigen/HLA/

	>>>
	output{
        String hla = "${outputDir}/neoantigen/HLA/${normal}_result.tsv"
    }
}

task neoantigen{
	String codes_dir
	String outputDir
	String name
	String normal
	String tumor
	String hla
	Array[String] vcf 
	command <<<
		sh /home/jm001/test_duantao/database_update/test_project/t20230818/predict_neoantigen.sh ${outputDir} ${name} ${tumor} ${codes_dir}
	>>>
	output{
        String neoantigen = "${outputDir}/neoantigen/MHC_Class_I/${tumor}.all_epitopes.netchop.txt"
    }
}

task auto_report{
	String cancer
	String codes_dir
	String outputDir
	String name
	String normal
	String tumor
	Array[String] qc_2
	Array[String] concordance
#	String purity
	String snv_result
	String cnv_result
	String fusion_result	
	String tmb

	command <<<
		
		perl /home/jm001/test_duantao/database_update/codes/682/indication.pl ${outputDir} ${cancer}
		python3 ${codes_dir}/qc_check_tissue.py ${outputDir} ${tumor} ${normal} ${name}
		python3 ${codes_dir}/drug_dedup.py ${outputDir} ${tumor}
        	perl ${codes_dir}/hpd.pl ${outputDir} ${tumor}
       		python3 ${codes_dir}/682gene_tissue_control_report.py ${outputDir} ${tumor} ${normal} ${cancer}
		python3 ${codes_dir}/wdl_check.py -o ${outputDir}
		perl ${codes_dir}/file_format_change.pl ${outputDir} ${tumor}
#		cp ${outputDir}/qc/sequenza/${name}_confints_CP.txt ${outputDir}/report/${name}_confints_CP.txt
		cp ${outputDir}/qc/qc_fail.txt ${outputDir}/report/qc_fail.txt
		cp ${outputDir}/qc/${tumor}_qc.txt ${outputDir}/report/
		cp ${outputDir}/cnvkit/${tumor}.rmdup.cns.cn ${outputDir}/report/
		cp ${outputDir}/cnvkit/${tumor}.cnv.png ${outputDir}/report/
		cp ${outputDir}/fusion/${tumor}.fuison.vus.txt ${outputDir}/report/
		cp ${outputDir}/fusion/${tumor}.fusion.reanno.vcf ${outputDir}/report/
#		cp ${outputDir}/mutation/*filter* ${outputDir}/report/
		cp ${outputDir}/mutation/${tumor}.snvindel.Germline.target.txt ${outputDir}/report/
		cp ${outputDir}/mutation/${tumor}.nontarget.vus.txt ${outputDir}/report/
		cp ${outputDir}/mutation/${tumor}.snvindel.vus.txt ${outputDir}/report/
		cp ${outputDir}/mutation/${tumor}.target.splicing.txt ${outputDir}/report/
		cp ${outputDir}/mutation/${tumor}.target.promoter.txt ${outputDir}/report/
		cp ${outputDir}/mutation/${tumor}.tmb.txt ${outputDir}/report/
		cp ${outputDir}/MMR/${tumor}.mmr.pre.txt ${outputDir}/report/  
		cp ${outputDir}/HRR/${tumor}.hrr.pre.txt ${outputDir}/report/
		cp ${outputDir}/HPD/${tumor}.hpd.pre.txt ${outputDir}/report/
		cp ${outputDir}/hereditary/${tumor}.hereditary.pre.txt ${outputDir}/report/
		python3 ${codes_dir}/txt_xlsx.py ${outputDir} ${tumor}
	>>>
}


# hotspot
task hotspot{
        String tumor
        String outputDir
        String ref
        Array[String] rmdupBam
        String codes_dir
        command<<<
        
        mkdir -p ${outputDir}/mutation/hotspot
        samtools mpileup -Bq 20 -Q 20  -f ${ref} -l ${codes_dir}/hotspot.bed  ${outputDir}/alignment/${tumor}.rmdup.bam -o ${outputDir}/mutation/hotspot/${tumor}.hotspot.pileup
        java -jar $VARSCAN mpileup2cns ${outputDir}/mutation/hotspot/${tumor}.hotspot.pileup --min-var-freq 0.005 --min-avg-qual 20 --output-vcf 1 --variants 1 --p-value 0.99 --min-reads2 2 --strand-filter 0 > ${outputDir}/mutation/hotspot/${tumor}.hotspot.L.snp.indel.vcf
        java -jar $VARSCAN mpileup2cns ${outputDir}/mutation/hotspot/${tumor}.hotspot.pileup --min-var-freq 0.01 --min-avg-qual 20 --output-vcf 1 --variants 1 --p-value 0.05 --min-reads2 3 --strand-filter 1 > ${outputDir}/mutation/hotspot/${tumor}.hotspot.H.snp.indel.vcf
        perl ${codes_dir}/hotspot.hvl.pl ${outputDir} ${tumor} 

        if [ -e "${outputDir}/mutation/hotspot/${tumor}.hotspot.snp.indel.vcf" ]; then
                table_annovar.pl \
                ${outputDir}/mutation/hotspot/${tumor}.hotspot.snp.indel.vcf \
                /dataseq/jmdna/software/annovar/humandb/ \
                -buildver hg19 -nastring . -vcfinput -remove -otherinfo \
                -protocol refGene \
		-argument '-hgvs' \
                -operation g \
                --outfile ${outputDir}/mutation/hotspot/${tumor}.hotspot.snp.indel.anno


#               java -jar /dataseq/jmdna/software/GenomeAnalysisTK.3.7.jar -T VariantAnnotator \
#               -R ${ref} \
#               -I ${outputDir}/alignment/${tumor}.rmdup.bam \
#               -V ${outputDir}/mutation/hotspot/${tumor}.hotspot.snp.indel.vcf \
#               -o ${outputDir}/mutation/hotspot/${tumor}.hotspot.TandemRepeatAnnotator.vcf \
#               --annotation TandemRepeatAnnotator

#               grep -v "^##" ${outputDir}/mutation/hotspot/${tumor}.hotspot.TandemRepeatAnnotator.vcf | cut -f8| paste ${outputDir}/mutation/hotspot/${tumor}.hotspot.snp.indel.anno.hg19_multianno.txt  -  >${outputDir}/mutation/hotspot/${tumor}.hotspot.snp.indel.Somatic.annoall.hg19_multianno.txt               
#        cp ${outputDir}/mutation/hotspot/${tumor}.hotspot.snp.indel.anno.hg19_multianno.txt ${outputDir}/report/${tumor}.hotspot.snp.indel.anno.hg19_multianno.txt
		
		perl ${codes_dir}/hotspot.filter.pl ${outputDir} ${tumor}
        fi
        
        >>>
        output{
                                String  hotspot = "${outputDir}/mutation/hotspot/${tumor}.hotspot.H.snp.indel.vcf"
        }
}