QAseq-Fusion_v2.1/py_script/breakpoint_stat_single_v6.1.py

#!/usr/bin/python
# -*- coding:utf-8 -*-

import argparse,time

gene_info = {}
gene_strand = {}

DB_DIR="/cgdata/IVDRD/18gene_DB/qaseq_fusion"

primer_homo_file=DB_DIR+"/sample_primer_stat.txt"
primer_seq_file=DB_DIR+"/select_primer_info_total.txt"
ref=DB_DIR+"/hg38.fa"
db=DB_DIR+"/hg38"
transcript_nm=DB_DIR+"/big_panel_transcript_nm_hg38.xls"


PRIMER_INFO={}
PRIMER_ID={}
PRIMER_HOMO={}
BASE_DICT={'A':'T','T':'A','G':'C','C':'G','N':'N'}

def get_primer_info():
    #PRIMER HOMO
    #{'primer':['primer1','primer2','primer3',...],}
    with open(primer_homo_file,'r') as phf:
        for line in phf:
            info = line.strip("\n").split("\t")
            PRIMER_HOMO[info[0]]=info[1].split(",")

    #PRIMER ID
    #{'primer':'ATGC','primer1':'ATGCCC'}
    #PRIMER INFO
    with open(primer_seq_file,'r') as psf:
        for line in psf:
            info = line.strip("\n").split("\t")
            PRIMER_ID[info[0]]=info[1]
            PRIMER_INFO[info[0]]=info

def get_PrimerPre_Site(PrimerPre_Site_Info_file,reads_Primer_dict):
    Info_dict={}
    with open(PrimerPre_Site_Info_file,'r') as PPSI:
        for line in PPSI:
            info = line.strip("\n").split("\t")
            ID=info[-1].split("_")[0]
            if ID == "":
                continue
            primerID=reads_Primer_dict[ID][0]
            if 'IPC' in primerID:
                continue
            
            mismatch_seq=info[2]
            match_seq=info[3]
            if info[1] =="Down":
                primerID=reads_Primer_dict[ID][1]
                if 'IPC' in primerID:
                    continue
                mismatch_seq="".join([BASE_DICT[base] for base in mismatch_seq][::-1])
                match_seq="".join([BASE_DICT[base] for base in match_seq][::-1])
            Info_dict[info[0]]=[info[0],primerID,mismatch_seq,match_seq,info[-2]]

    return Info_dict

def out_test(dict):
    for key in dict.keys():
        print("TEST:")
        print(key)
        print(dict[key])

def OvarLap(seq1,seq2,MISMATCH=1):
    num = len(seq1)
    count=0
    for index in range(num):
        if seq1[index] !=seq2[index]:
            count +=1
        if count >MISMATCH:
            return False
    return True

def check(primer_seq,primer_down_seq,mismatch_seq):
    mismatch_len=len(mismatch_seq)
    print("TEST")
    print(primer_seq)
    print(primer_down_seq)
    print(mismatch_seq)
    for index in range(mismatch_len,5,-1):
        num=min(len(primer_seq),mismatch_len-index)
        for match_index in range(num,5,-1):
            primer_temp_seq=primer_seq[-1*match_index:]+primer_down_seq[:index]
            mismatch_temp_seq=mismatch_seq[-1*index-match_index:]
            if OvarLap(primer_temp_seq,mismatch_temp_seq):
                return True,index,match_index+index
    return False,0,0

def get_primer_Site(primer_list,mismatch_seq,match_seq):

    primer_candidate_fusion_dict={}
    for primer in primer_list:
        primer_temp=PRIMER_INFO[primer]
        primer_down_seq=primer_temp[-1]
        primer_up_seq=primer_temp[-2]
        primer_seq=primer_temp[1]
        check_results=check(primer_seq,primer_down_seq,mismatch_seq)
        print(check_results)
        if check_results[0]:
            match_common_len=int(check_results[1])
            total_match_common_len=int(check_results[2])
            strand = primer_temp[5]
            primer_chr=primer_temp[2]
            primer_start=int(primer_temp[3])
            primer_end=int(primer_temp[4])
            breakpoints=primer_end+match_common_len
            site_e=primer_chr+":"+str(breakpoints-30)
            if match_common_len>=30:
                fusion_seq=primer_down_seq[match_common_len-30:match_common_len]+match_seq[:30]
            else:
                if match_common_len+len(primer_seq)>=30:
                    fusion_seq=primer_seq[-1*(30-match_common_len):]+primer_down_seq[:match_common_len]+match_seq[:30]
                elif match_common_len+len(primer_seq)>=15:
                    primer_seq_len=len(primer_seq)
                    fusion_seq=primer_up_seq[-1*(30-match_common_len-primer_seq_len):]+primer_seq+primer_down_seq[:match_common_len]+match_seq[:30]
            if strand == '-1':
                breakpoints=primer_start-match_common_len
                site_e=primer_chr+":"+str(breakpoints+30)
            site=primer_chr+":"+str(breakpoints)
            primer_candidate_fusion_dict[primer]=[primer,site,site_e,fusion_seq,str(total_match_common_len)]
    return primer_candidate_fusion_dict

def get_Uniq_UMIs(ID_list):
    UMIs_dict={}
    for ID in ID_list:
        UMI=ID.split("_")[2]
        UMIs_dict[UMI]=0
    
    return len(UMIs_dict)

def get_single_primer_points(PrimerPre_Site_Info,indir):

    #记录每个readsID 对应site
    single_points_PrimersPre_readsInfo_file=indir+"/single_points_PrimersPre_readsInfo.txt"
    single_points_PrimersPre_readsInfo_dict={}
    with open(single_points_PrimersPre_readsInfo_file,'r') as sppr:
        for line in sppr:
            info=line.strip("\n").split("\t")
            if info[0] not in single_points_PrimersPre_readsInfo_dict.keys():
                single_points_PrimersPre_readsInfo_dict[info[0]]=[info[-1]]
            else:
                single_points_PrimersPre_readsInfo_dict[info[0]].append(info[-1])
 
    fusion_breakpoints_detail_file=indir+"/single_fusion_breakpoints_detail.txt"
    fusion_breakpoints_detail=open(fusion_breakpoints_detail_file,'w')
    fusion_breakpoints_detail.write("\t".join(['KeyPoints','Points','Point1_End','Point2_End','FusionSeq','Primer'])+"\n")

    fusion_breakpoints_readsID_file=indir+"/single_fusion_breakpoints_readsID.txt"
    fusion_breakpoints_readsID=open(fusion_breakpoints_readsID_file,'w')
    fusion_breakpoints_readsID.write("\t".join(['Points','FusionSeq','ReadsID'])+"\n")

    fusion_breakpoints_stat_file=indir+"/single_fusion_breakpoints_stat.txt"
    fusion_breakpoints_stat=open(fusion_breakpoints_stat_file,'w')
    fusion_breakpoints_stat.write("\t".join(['Points','Point1_End','Point2_End',"Overlap",'DoubleReads','SingleReads','UMIkinds','FusionSeq'])+"\n")

    for key_site in PrimerPre_Site_Info.keys():
        temp_info_dict=PrimerPre_Site_Info[key_site]
        primerID=temp_info_dict[1]
        key_site_end=temp_info_dict[-1]
        if primerID in PRIMER_HOMO.keys():
            primer_list=PRIMER_HOMO[primerID]
        else:
            primer_list=[]
        primer_list.append(primerID)
        
        primer_candidate_fusion_dict=get_primer_Site(primer_list,temp_info_dict[2],temp_info_dict[3])
       
        for item in primer_candidate_fusion_dict.keys():
            item_temp=primer_candidate_fusion_dict[item]
            site=item_temp[1]+"-"+key_site
            fusion_breakpoints_detail.write("\t".join([key_site,site,item_temp[2],key_site_end,item_temp[3],item])+"\n")
            
            ID_list=single_points_PrimersPre_readsInfo_dict[key_site]
            SingleID=len(ID_list)
            UMIkinds=get_Uniq_UMIs(ID_list)
            fusion_breakpoints_stat.write("\t".join([site,item_temp[2],key_site_end,str(0),str(0),str(SingleID),str(UMIkinds),item_temp[3]])+"\n")
            for ID in ID_list:
                fusion_breakpoints_readsID.write("\t".join([site,item_temp[-2],ID])+"\n")

    fusion_breakpoints_detail.close()
    fusion_breakpoints_readsID.close()
    fusion_breakpoints_stat.close()

def get_reads_primers_info(readsPrimer):
    
    reads_Primer_dict={}
    with open(readsPrimer,'r') as rp:
        for line in rp:
            info=line.strip("\n").split("\t")
            if len(info)==10:
                reads_Primer_dict[info[0]]=[info[3],info[7]]
    return reads_Primer_dict

def main(indir,readsPrimer):

    t1=time.time()
    get_primer_info()

    #读取reads primer信息
    reads_Primer_dict=get_reads_primers_info(readsPrimer)

    PrimerPre_Site_Info_file=indir+"/single_points_PrimersPre_site.txt"
    PrimerPre_Site_Info=get_PrimerPre_Site(PrimerPre_Site_Info_file,reads_Primer_dict)
    print(PrimerPre_Site_Info)
    get_single_primer_points(PrimerPre_Site_Info,indir)

    t2=time.time()
    print("Times: "+str(int(t2-t1))+"s")
    print("BreakPoints Stat Single Done!")


if __name__ == '__main__':
    parser = argparse.ArgumentParser(description='single breakpoints primers')
    parser.add_argument('-i', required=True, type=str, help="indir")
    parser.add_argument('-r', required=True, type=str, help="reads primers file")
    args = parser.parse_args()
    main(args.i,args.r)