ABSTRACT
Objective: Based on optimized method of DNA ligase chain reaction in medium/low throughput genotyping, we assessed the relationship between NOD-like receptor genes NOD1, NOD2 and coronary artery disease (CAD) occurrence. Methods: A multiplex PCR was conducted to enrich DNA template; probe design, annealing temperature, time and number of circulation of PCR were opfimizecl for allele specific ligation; allele specific products were identified by fluorescence PCR and capillary electrophoresis; the accuracy was verified by Sanger sequencing. Single nucleotide polymorphisms (SNPs) on NOD1 gene and NOD2 gene were examined in 1555 CAD patients and 1887 control subjects; the relationship between SNPs and CAD occurrence was studied. Results: Based on optimized PCR condition and allele specific probe design, 30 allele loci genotyping can be genotyped by 10ng DNA template at one time. Association study presented that rs751271 and rs1861759 on NOD2 gene were related to non-hypertensive CAD, allP<0.05; with Bonferroni correction, such correlation was still significant, allP<0.05. Conclusion: We optimized DNA ligase chain reaction and established a novel high accuracy, low cost method for the demand of medium/low throughput genotyping in clinical molecular diagnosis. With this method, we identified that rs1861759 and rs751271 on NOD2 gene were associated with non-hypertensive CAD.
ABSTRACT
BACKGROUND: Although it is known that patients wth peripheral vascular disease are at high risk for coronary arterial disease, however, it has not been determined that patients with coronary artery disease(CAD) correlates with peripheral vascular disease(PVD). This study was designed to determine the prevalence and clinical characteristics of peripheral vascular disease(PVD) in patients with coronary artery disease(CAD). METHODS: A total of one hundred seventy-eight patients with CAD confirmed by coronary angiogram(145 male, age 58.5+/-10.1) were included in this study from February 1992 to May 1994. Coronary and peripheral angiograms were performed in all patients and the patients were divided into two groups; patients with PVD dand patients without PVD. Clinical characteristics were compared between two groups. RESULTS: Peripheral vascular disease was present in 49 patients(27.5%) among 178 CAD patients. The mean age of patients with PVD was significantly older than that of patients without PVD. The hypertension was statistically significant difference between two groups(P<0.05). There were no major differences in the number of risk factors or number of stenotic coronary arteries in patients with and without PVD. CONCLUSION: The prevalence of PVD in patients with CAD was high and it is reasonable to state that most common risk factors for coronary and peripheral atherosclerosis were age, hypertension and obesity.