Establishment Of Molecular Diagnosis Method For Gout-Related Gene SNPs And Comparative Evaluation

Objectives:To establish a matrix-assisted laser desorption ionization time-of-flight mass spectrometry(MALDI-TOF)molecular diagnostic method for gout-related genes single nucleotide polymorphism(SNP)loci and complete the genotyping of 48 SNP loci.To validate relationship of those loci with the susceptibility of gout populations in Northwestern China.To make a methodological evaluation of MALDI-TOF and PCRHRM.Methods:We designed specific amplification primers and single base extension primers of 48 SNP sites by software and established a molecular diagnostic method based on MALDI-TOF technology.The genotypes and allele frequencies of the case group and the control group were analyzed by statistical software.The results of two methods were compared and validated by Sanger sequencing to evaluate the detection performance.Results:(1)The established MALDI-TOF detection system completed the detection of 48 SNP loci in 400 samples,with an overall call frequency of 99.73%(19148/19200),and 52 genotypes of 7 loci failed to genotype.The allele of 19 SNPs in 12 genes may increase the risk of gout,and other 19 SNPs allele in 11 genes may lower the risk of onset of gout.(2)The coincidence rate between the genotyping results of two methods is 96.27%,and the consistent results is 100% correct when compared with Sanger sequencing.Conclusion:The MALDI-TOF molecular diagnostic method established in this study can perform high-throughput and rapid genotyping with a high call frequency.ABCG2,APOA4,APOBEC4,TNRC6 C,PTGR2,PKD2,ZNF410,FAM161 B,EXOC3L2,TMC6,RGS20,SLC22A12,ENTPD5,GCKR,SLC2A9,and SLC17A1 gene polymorphisms are related to the genetic susceptibility to gout in Northwestern China.The MALDI-TOF method is suitable for the scientific research of multiple sites in a large sample of a population,and the PCR-HRM method is suitable for conventional laboratories for scientific research and clinical diagnosis.