Establishment Of ALDH2 Glu504Lys Gene Polymorphism Detection Method And Its Correlation With Coronary Atherosclerotic Heart Disease

Coronary atherosclerotic heart disease(CAD)is the main cause of human death and disability worldwide.China is a major country suffering from CAD,and its prevalence rate is rising continuously.As a result,the direct economic burden caused by it will also increase.Therefore,the early diagnosis and prevention of the occurrence of CAD is very important.Nitroglycerin is the main treatment for CAD,ALDH2 is a key enzyme of nitroglycerin effective metabolite of nitric oxide,and its genetic polymorphism directly affects the efficacy of nitroglycerin in patients with angina pectoris.On the other hand,the genome-wide association analysis found that the polymorphism of the ALDH2 Glu504 lys gene(also known as rs671)was associated with CAD.However,due to the obvious ethnic and regional differences in the distribution of ALDH2 gene rs671 and the number of cases is limited,the association between ALDH2 gene rs671 and CAD is still controversial in China.At present,detection methods for the ALDH2 gene rs671 are expensive or require complicated procedural requirements.Therefore,the establishment of a simple,rapid and economical detection method for the ALDH2 gene rs671 loci not only provides a reference basis for the rational use of nitroglycerin in patients with angina pectoris,but also collect data on the correlational research between rs671 and CAD in different regions in China.This study established a convenient,accurate and economical detection method for the detection of ALDH2 gene rs671 locus.Using Sanger sequencing as a control method,Totally 1018 clinical samples were performed using this method in Beijing,Hunan and Guangzhou.At the same time,838 cases of clinical samples were analyzed for the association of genetic polymorphisms and CAD.The main research results are as follows:1.Based on the GoldMag nanoparticle chromatography system combined with ARMS-PCR principle,an ARMS-PCR GoldMag chromatography for the ALDH2 gene rs671 loci was successfully established.The sensitivity,accuracy,and batch-to-batch performance of the method were evaluated and verified by clinical trials.It was proved that the method results was completely consistent with the traditional sequencing method.The coincidence rate was 100%,and it has the advantages of cheap,simple,fast,high sensitivity,reliability,wide applicability and so on.2.The ALDH2 gene polymorphisms in Beijing,Hunan,and Guangzhou were detectedand statistically analyzed using established methods.It was found that the mutation frequency of the ALDH2 gene rs671 locus in Beijing was low,and the frequency of the mutation allele A was only 10.9%;Hunan and Guangzhou the distribution of ALDH2 gene polymorphisms was similar,with higher mutation frequencies than in Beijing.It shows that the diversity of ALDH2 gene distribution in different regions of China does exist.3.Through a case-control study,the association between ALDH2 gene polymorphisms and CAD in northern China(Beijing)and southern China(Guangzhou and Hunan)was analyzed.It was found that ALDH2 gene polymorphisms in Beijing were associated with CAD.The frequency of mutations in the CAD groups was significantly higher the control group;however,the same analysis of the Hunan and Guangzhou regions did not find this correlation.The difference of these results further illustrates the necessity of studying the correlation between rs671 and CAD in different regions of China,thus providing clinical reference significance for the prevention of coronary heart disease in different regions and the use of nitroglycerin in patients.