Genotyping Of Candidate Genes Based On GoldMag Lateral Flow Assay For Assessing Essential Hypertension Risk

Essential hypertension is a polygenetic disease affected by genetic and environmental factors,and the study of related or susceptible genes to the disease has attracted lots of attention.The detection of essential hypertension susceptibility genes can especially help in finding potential risk of disease,which benefit for the early treatment and reducing the risk of cardiovascular and cerebrovascular diseases(such as stroke)of patients with essential hypertension.Current genotyping method include PCR-RFLP,fluorescence PCR,gene sequencing and gene chip that need complicated operation steps or special instruments and have to be carried out for a long period.In this study,ACE(I/D)and AGTR1(A1166C),two candidate susceptibility genes of essential hypertension,were selected as the research objects.A rapid genotyping method called T-ARMS-LFA was established by combining tetra-primer amplification refractory mutation system polymerase chain reaction(tetra-primer ARMS-PCR)and Gold Mag lateral flow Assay(LFA).A single nucleotide polymorphism(AGTR1)or deletion(ACE)of a gene can be accomplished by employing four primers in one PCR reaction and interpreting result via lateral flow strip.Compared with the gene sequencing method,the detection system present coincidence rate of 100%.The sensitivity for genotyping of ACE(I/D)and AGTR1(A1166C)were 25 ng and 10 ng respectively.The genotypes of ACE(I/D)and AGTR1(A1166C)in 135 cases with essential hypertension and 189 healthy controls of Northwestern China were analyzed by T-ARMS-LFA.The statistical analysis results showed that the genotypes of ACE(I/D)and AGTR1(A1166C)in the experimental group of essential hypertension and the control group of healthy people were consistent with Hardy Weinberg equilibrium.It was found that there was a significant correlation between the D allele of ACE gene and essential hypertension in the log additive model(OR = 1.51,95% CI = 1.02-2.23,p = 0.039),which provided a clinical reference for the risk of essential hypertension.The genotyping method established in this research was simple,accuracy and stable,which is significant to large-scale universal screening in hypertension population for chronic disease management.