Study On The Correlation Of CBS C.16153 C>T With Serum Homocysteine Content And The Establishment Of HRM Detection Method In Wannan Area

Objective:1.Establishment of high-resolution melting detection method for SNP site of CBS.2.SNP sites of CBS c.16153 C>T in high homocysteine population in Wannan area.Methods:1.Search for key enzymes in the homocysteine metabolic pathway at the Pub Med website.2.The gene sequences of key enzymes were queried on NCBI website,and primers were designed for common mutation sites.3.Standard samples were collected and peripheral blood mononuclear cells were extracted.The collected PBMC were used for DNA extraction,storage and reserve.4.The designed primers and extracted DNA were amplified by PCR to explore the optimal procedure and concentration of the experiment.5.The primers and DNA samples were amplified by PCR to explore the optimal procedure and concentration of the experiment.6.The PCR products were analyzed by high-resolution melting curve.7.The results of high resolution melting curve and sequencing were analyzed.8.Select SNP sites successfully established for detection of HRM technology and continue the experiment to analyze the relationship between SNP sites and HCY content.8.Combined with clinical data,Chi-square test and two independent sample test were used for statistical analysisResults:1.The rate of CBS c.16153 C>T site of CBS was statistically significant in normal people and patients with high homocysteine(P <0.05).2.In the normal group and all subjects,the HCY content of specimens containing site 16153 mutation was statistically significant compared with that of samples without mutation(P <0.05).Conclusion:1.The detection method of CBS c.16153 C>T high resolution melting curve was successfully established.2.In the study subjects,the mutation rate of CBSc.16153 C>T site was significantly higher i n normal people than in patients with high homocysteine.3.In the normal group and the ove rall study object,the HCY content of specimens containing site 16153 mutation was significantly lower than that of specimens without mutation.