Mutation Analysis And Pathogenic Gene Localization Of Candidate Genes In A Benign Familial Infantile Epilepsy Family

Objective: The purpose of this study was to locate a disease-causing gene in a benign familial infantile epilepsy family at the epilepsy center of the Second Hospital of Lanzhou.Using a method of linkage analysis and whole exon sequencing by extracting clinical phenotypes and biological samples.The pathogenic genes and genetic polymorphisms of the family are found,thereby locating the pathogenic genes of the family,suggesting the pathophysiological molecular mechanism,providing a theoretical basis for the development of genome therapy and new drugs,and improving the targeting of the treatment.Methods: This study collected a benign familial infantile epilepsy family with 12 members,including 5 patients and 7 non-patients.The above-mentioned family members were collected 5 ml of peripheral blood and extracted g DNA.Based on the results of epileptic gene package of 551 epileptic genes,two genes,PRRT2 gene and RHAG gene were screened,and one generation of sequencing was verified.The sequence was from NCBI and the total volume of the PCR reaction was 50 ul.The size of the amplification results was determined by agarose gel electrophoresis.The linkage analysis uses LINKAGE software to determine whether the linkage is based on the LOD value in the autosomal dominant inheritance mode.DNA from the entire exon region was enriched using Agilent's LC chip capture system and then subjected to high-throughput sequencing on the Illumina Hi Seq 2500/4000 sequencing platform.The db SNP database,1000 d Genome database,SIFT and Poly Phen2 software were used to screen the genes.Results: The Sanger sequencing results of PRRT2 gene and RHAG gene did not conform to Mendelian inheritance law.The LOD value of family linkage analysis was between 1-1.5,and WES sequencing results screened 29 candidate genes.Conclusion: The PRRT2 gene is not the causative gene of the BFIE family;the RHAG gene is not the causative gene of benign familial infantile epilepsy;the linkage analysis of the family does not identify the chromosomal region with LOD value greater than 3.The results of the whole exon sequencing screened out 29 candidate genes.