#!/usr/bin/env bash
##Author: guochangquan
##Copyright 20210929
##company: NuProbe

set -e

function usage() {
    echo "Usage: $0 [ -s <sample> ] [ -i <fq_dir> ] [ -o <analysis_dir> ] [ -x <script_dir> ] [ -c <FeBY2_CNV_outdir> ]"
    echo "-s sample name"
    echo "-i input fastq dir"
    echo "-x script_dir"
    echo "-o analysis_dir"
    echo "-c FeBY2 CNV outdir"
}

if [ $# -eq 0  ]
then
    usage
    exit
fi

while getopts ":s:c:x:o:i:h" opt; do
    case $opt in
        i)
            fq_dir=$OPTARG
            ;;
        o)
            out_dir=$OPTARG
            ;;
        s)
            sample=$OPTARG
            ;;
        c)
            cnv_dir=$OPTARG
            ;;
        x)
            feby2_script=$OPTARG
            ;;
        h)
            usage
            exit
            ;;
        \?)
            echo "Invalid option: -$OPTARG" > /dev/stderr
            usage
            exit
            ;;
    esac
done
echo "Input FASTQ dir:" $fq_dir
echo "Output BAM dir:" $out_dir
echo "Sample Name:" $sample
echo "CNV Analysis dir:" $cnv_dir
echo $feby2_script

. $feby2_script/conf/feby2.cfg

### FASTQ TRIM and Mapping
test -d $out_dir/${sample} || mkdir ${outdir}/${sample}
cd $out_dir/${sample}
$fastp -i ${fq_dir}/${sample}/${sample}_combined_R1.fastq.gz -I ${fq_dir}/${sample}/${sample}_combined_R2.fastq.gz -o ${sample}_1.fq.gz -O ${sample}_2.fq.gz -c --length_required 50 -q 20 -j ${sample}.fastp.json -h ${sample}.fastp.html
$speedseq_dir/speedseq align -o ${sample} -M 20 -t 8 -R "@RG\tID:${sample}\tSM:${sample}\tLB:${sample}\tPL:ILLUMINA" $ref ${sample}_1.fq.gz ${sample}_2.fq.gz && rm ${sample}_1.fq.gz ${sample}_2.fq.gz

### SNV/INDEL CAll and annotation
$gatk4 --java-options "-Xmx10g" HaplotypeCaller -R $ref -I ${sample}.bam -ip 20 -L $target_call -O ${sample}.gatk.vcf.gz
/cgdata/CGTools/CGTools/wrapper/S14-annovar-snv-annotation.sh -w . -s  ${sample}.gatk  -v ${sample}.gatk.vcf.gz
perl $exe_dir/snp_stat.pl $sample.gatk.hg19_multianno.txt
python /cgdata/CGTools/CGTools/tools/T07-var-filter.py -i $sample.gatk.hg19_multianno.txt -o $sample.gatk.hg19_multianno.xlsx

### QC
test -d conest/ || mkdir conest/
$qualimap_dir/qualimap bamqc -bam ${sample}.bam -c --java-mem-size=20G -sd -gff ${target}
$verify --vcf $bia_snp --bam ${sample}.bam --out conest/${sample}.verify --verbose --ignoreRG &
$gatk4 CollectHsMetrics -I ${sample}.bam -BI $interval -TI $interval -O ${sample}_hs_metrics.txt -R $ref --COVERAGE_CAP 6000 -MQ 0 --CLIP_OVERLAPPING_READS false &
$gatk4 GetPileupSummaries -I ${sample}.bam -L $target -V $bia_snp -max-af 0.99 -min-af 0.01 -O conest/${sample}.ps.tab
$gatk4 CalculateContamination -I conest/${sample}.ps.tab -O conest/${sample}.contEST.tab
wait

### yesuan
$gatk4 HaplotypeCaller -R $ref -I ${sample}.bam --genotyping-mode GENOTYPE_GIVEN_ALLELES -L $ye_target --alleles $ye_site -O ${sample}.ye.vcf
bcftools norm -m-both -o ${sample}.step1.vcf ${sample}.ye.vcf
bcftools norm -f $ref -o ${sample}.step2.vcf ${sample}.step1.vcf
$gatk4 VariantsToTable -V ${sample}.step2.vcf -O ${sample}.ye.norm.tab -F CHROM -F POS -F TYPE -GF GT -GF GQ -GF AD -F REF -F ALT --show-filtered && rm ${sample}.step2.vcf ${sample}.step1.vcf
perl $ye_home/prep_FeBY2_plus.pl ${sample} FeBY2 $ye_ref

### prepare for CNV
bamf=$out_dir/$sample/$sample.bam
cd $cnv_dir
samtools view -f 2 -F 1024 -F 2048 -hub $bamf  \
| samtools sort -n -l 0 -m 20G - |bamToBed -bedpe \
| awk '$1!~/M/ && $1==$4 && $2!=-1 && $6-$2>0 && $6-$2<1000{print $1"\t"$2"\t"$6"\t"$7}' \
| intersectBed -a - -b $cnv_target -loj -wb \
| perl $feby2_script/script/get_GCcor2cov.pl $sample $ref $target_GCref $cnv_target b37 > $sample.gc.xls

test -s $sample_?_QC_plot.pdf || Rscript $feby2_script/script/MBY2_CNV_stat.r $sample 
samtools view -hb -L $target $bamf > ${sample}.target.bam && samtools index ${sample}.target.bam && samtools idxstats ${sample}.target.bam > ${sample}.idxstats && rm ${sample}.target.bam*