Based On DNA-pooling Genome-wide Sequencing For Primary Screening Of Coronary Atherosclerotic Heart Disease Susceptibility Gene Mutation

OBJECTIVE: Using DNApooling genome heavy sequencing technology to screen the CAD susceptibility gene mutation sites.MEHODS: 100 cases of blood samples from CAD group and control group were collected.DNA is extracted from blood cells to make DNApooling for sequencing.The sequencing results of the two groups were compared with NCBI human gene pool hg19,and the mutations were selected and the ANNOVAR software was used to classify all the mutations.The metabolic analysis of the two groups was analyzed by Fisher Exact Test.Detection of significant(p <0.01)sites that are estimated to be associated with the disease-related mutations.The mutations were analyzed by Gene Ontology analysis to screen these gene mutation sites for preliminary functional annotation.RESULTS:(1)CAD groups DNA sequencing generated 1057326436 raw reads based of data yielding 32.76 x sequencing depth.99.84% were successfully mapped to the reference genome by the BWA mapping approach.Control groups DNA sequencing generated 1016731048 raw reads based of data yielding 36.13 x sequencing depth.99.88% were successfully mapped to the reference genome by BWA mapping approach.The CAD group were matched with the reference reads by994931429(95.62%)and the control is 956586404(95.68%).The error matching rates were 0.57% and 0.57% of the CAD group and the control group.(2)We received a total of 28,772 SNPs loci,17,654 InDels sites,278 SVs sites,882 CNVs loci by analyzing data of obtained earlier.We found that the C:G?T:A motation type is the most variation type in genome-wide and coding region.We got12,314 and 91 sites information respectively.(3)The protein-protein interaction(PPI)network was constructed by cytoscape software,and we finally get 73 susceptible genes susceptible to the CAD which are SERPINA3,ABL,ADRA1 A,FAS,ATBF1,BDKRB2,BSG,SERPING1,C3,DDR1,CASR,CD4,CD9,COL4A3,COL5A1,CP,CR1,CSNK2 B,CSPG2,EPHB1,PTK2 B,FBLN2,FBN2,GLG1,GNA12,GNAS,GRM5,HD,HLA-A HLA-B,HLA-DQA1,HLA-DQA2,HLA-DQB1,HLA-DRA,IGF1 R,RBPSUH,IL1 A,IL1RN,IL4 R,ITGA2B,ITGB3,KNG1,LCT,LRP2,MAPT,MSR1,NFKB1,P2RX7,PLCG2,PIK8,MAPK9,MASP1,RAF1,SELL,TG,TGFBR2,TNR,USH2 A,LRP8,CUBN,RAPGEF4,AKAP13,CARD8,MACF1,CABIN1,HIPK2 and MUC16.(4)The function of the mutated gene was analyzed by Gene Ontology analysis function software.It was found that the mutated genes were mainly involved in inflammatory cytokine regulation,lipid metabolism regulation,coagulation and fibrinolysis,and bioadhesion.CONCLUSIONS:(1)The CAD susceptibility genes were screened at the whole genome level by genome-wide re-sequencing,and 73 CAD-related susceptibility gene mutations were screened by PPI network analysis:SERPINA3,ABL,ADRA1 A,FAS,ATBF1,BDKRB2,BSG,SERPING1,C3,DDR1,CASR,CD4,CD9,COL4A3,COL5A1,CP,CR1,CSNK2 B,CSPG2,EPHB1,PTK2 B,FBLN2,FBN2,GLG1,GNA12,GNAS,GRM5,HD,HLA-A HLA-B,HLA-DQA1,HLA-DQA2,HLA-DQB1,HLA-DRA,IGF1 R,RBPSUH,IL1 A,IL1RN,IL4 R,ITGA2B,ITGB3,KNG1,LCT,LRP2,MAPT,MSR1,NFKB1,P2RX7,PLCG2,PIK8,MAPK9,MASP1,RAF1,SELL,TG,TGFBR2,TNR,USH2 A,LRP8,CUBN,RAPGEF4,AKAP13,CARD8,MACF1,CABIN1,HIPK2 and MUC16.(2)The function of the mutated gene was analyzed by Gene Ontology analysis function software.It was found that the mutated genes were mainly involved in inflammatory cytokine regulation,lipid metabolism regulation,coagulation and fibrinolysis,and bioadhesion.(3)The application of DNApooling technology to gene research for related diseases has greatly reduced the time and expense of research funding.