Research And Application Of Drug Related Gene Mutation Detection Method In Hypertension

As a new medical concept, personalized medicine is the trend of the development of clinical medicine in twenty-first Century. Before the patient use of drugs, detecting some drug related genes, not only can reduce the patient’s medical cost, improve drug efficacy, but also can avoid the occurrence of adverse drug reaction. In view of the above consideration, In this paper, with the aid of pyrophosphate sequencing platform and PCR-electrophoresis technology, the high blood pressure drug related genes and K-ras gene detection method research.the main work is as follows:The second chapter:ADRB1 gene is the targets of (31 adrenal receptor blockers. The mutant heterozygote (G1y389Arg) and mutant homozygous (Arg389Arg) of AD RBI gene is sensitive for β1 receptor blockers, but not wild homozygous (Gly389Gly). So the detection of ADRB1 gene may help hypertension patients to choose the right drugs. This paper adopted pyrosequencing technology to detect ADRB1 gene polymorphisms of hypertension patients, at the same time verified the methodology of pyrophosphate by capillary electrophoresis sequencing technology. Results show that the method detection accuracy is 100%. It can be used as a new method for the detection of ADRB1 Arg389G1y gene polymorphism in scientific research or clinical. Based on 184 cases of patients with high blood pressure,results show Gly389Gly genotype frequencies was 9.24%, Arg389Gly genotype frequencies was 38.04%, Arg389Arg genotype frequency was 52.72%.The third chapter:Cytochrome oxidase P450 2C9(CYP2C9)(Ile359>Leu359) mutation make CYP2C9 enzyme activity is differences between different dividuals. CYP2C9*3 mutation can reduce the blood drug concentration of E3174, which leads to weaken losartan antihypertensive effect. On the other hand CYP2C9*3 mutation can increase the blood drug concentration of valsartan and irbesartan, which leads to strengthen valsartan and irbesartan antihypertensive effect. So the detection of CYP2C9*3 gene polymorphism can guide hypertension patients to choose the appropriate drugs. This paper adopted pyro sequencing technology to detect CYP2C9*3 gene polymorphisms of hypertension patients, at the same time verified the methodology of pyrophosphate by capillary electrophoresis sequencing technology. Results show that the method detection accuracy is 100%. It can be used as a new method for the detection of CYP2C9*3 gene polymorphism in scientific research or clinical. Based on 184 cases of patients with high blood pressure,results show*1/*3 genotype frequencies was 7.6%,*1/*1 genotype frequencies was 92.4%,*3/*3 genotype did not appear.The fourth chapter:In the human body, angiotensin converting enzyme (ACE) level is controlled by ACE gene, so different ACE genotype differences in the levels of ACE is obvious. As ACE gene polymorphism have significant impact on antihypertensive efficacy of angiotensin converting enzyme inhibitor (ACEI), different ACE I/D genotype individuals have different sensitivity to certain ACEI drugs. In order to achieve the best effect, we should make patients’s ACE I/D genotype clear before choose drugs. This paper adopted PCR-electrophoresis analysis method to detect ACE I/D polymorphism of hypertension patients, and verified it by capillary electrophoresis sequencing technology. Results show that the method detection accuracy is 100%. It can be used as a new method for the detection of ACE I/D gene polymorphism in scientific research or clinical. Based on 184 cases of patients with high blood pressure,results show I/I genotype frequencies was 44.57%, I/D genotype frequencies was 40.76%, D/D genotype frequencies was 14.67%.The fifth Chapter:The epidermal growth factor receptor (EGFR) is the targets of Cetuximab. Patients with K-ras gene mutation use cetuximab will reduce the curative effect or even invalid. Most K-ras gene mutations occur in 12th codon[35G>A、35G>T、35G>C、 34G>A、34G>T、34G>C] and 13th codon [38G>A] of 1 exon (about 90%). In addition, about 5% of the mutation occur in the 61th codon of 2 exon. So the detection of K-ras gene mutation can predict the curative effect of Cetuximab.This paper adopted pyrosequencing technology to detect K-ras gene mutations, and verified it by the external quality assessment. Results show that the method detection accuracy is 100%. It can be used as a new method for the detection of K-ras gene mutation in scientific research or clinical. Based on 230 cases of patients with colorectal cancer,results show the K-ras gene mutation ratio was 14.34%. It was below the reported data of previous research results.