摘要
目的建立APOB基因rs693位点、LDLR基因rs688位点和APOC-Ⅰ基因rs4420638位点单核苷酸多态性（SNP）的聚合酶链式反应-高分辨率熔解曲线分析（PCR-HRM）的检测方法,并探讨该3个SNP位点与冠心病（CHD）易感的相关性。方法针对rs693、rs688和rs4420638 3个位点分别设计引物,建立PCR-HRM分子诊断方法,并通过直接测序进行验证。对311例CHD患者和300例体检健康者进行病例-对照研究,分析其与CHD易感性及血脂指标（TC、TG、HDL-C、LDL-C）的关系。结果rs693和rs4420638位点的基因频率和基因型频率在CHD组与对照组之间差异有统计学意义（P〈0.05）。rs693位点CT基因型降低CHD患病风险（OR=0.448,95%CI：0.246～0.817,P=0.009）;rs4420638位点AG基因型增加CHD患病风险（OR=2.140,95%CI：1.241～3.688,P=0.006）。rs688（C〉T）和rs4420638（A〉G）位点组成的单倍型CG增加CHD患病风险（OR=1.715,95%CI：1.091～2.697,P=0.018）。病例组rs4420638位点AG基因型患者的血清TC水平显著高于AA基因型（F=4.281,P=0.040）。结论成功建立了rs693、rs688和rs4420638 3个SNP位点的PCR-HRM基因分型方法。rs693和rs4420638位点以及单倍型CG与中国兰州地区汉族人群CHD易感性有关,rs4420638位点联合血清TC水平检测可预警CHD发生。
Objective To establish polymerase chain reaction and high resolution melting（ PCR-HRM） assays for genotyping three single nucleotide polymorphisms（ SNP）,i. e.,rs693 in apolipoproteins B（ APOB） gene,rs688 in low-density lipoprotein receptor（ LDLR） gene and rs4420638 in apolipoprotein C-Ⅰ（ APOC-Ι） gene,and investigate the relationship of the three SNPs with the risk of coronary heart disease（ CHD） development. Methods The molecular diagnosis methods of PCR-HRM were established by designing the primers of the three SNPs,rs693,rs688 and rs4420638. The accuracy of the developed methods was verified using direct sequencing of the amplicons. A case-control study was performed in 311 CHD patients and 300 matched healthy controls to analyze the correlation of the three SNPs with the susceptibility of CHD and blood lipid levels,total cholesterol（ TC）,triglyceride（ TG）,high density lipoprotein- cholesterol（ HDL-C）,and low density lipoprotein-cholesterol（ LDL-C） in CHD patients of Chinese Han population from Lanzhou region. Results The allele and genotype frequencies of rs693 and rs4420638 were significantly different between CHD cases and the controls（ P〈0. 05）. The genotype CT of rs693 confered to low risk of CHD development（ OR = 0. 448,95% CI： 0. 246-0. 817,P = 0. 009）,while the genotype AG of rs4420638 conferred to high risk of CHD（ OR = 2. 140,95% CI： 1. 241-3. 688,P =0. 006）. The haplotype analysis of the two SNPs（ rs688 and rs4420638） showed that haplotype CG increases the risk of CHD development（ OR = 1. 715,95% CI： 1. 091-2. 697,P = 0. 018）. The genotype AG of rs4420638 was significantly associated with the high TC level in CHD cases（ F = 4. 281,P = 0. 040）. Conclusion The present study suggested that the two SNPs（ rs693 and rs4420638）and the haplotype CG are significantly associated with the susceptibility of CHD development in Chinese Han population. The combined detections of rs4420638 and TC should be helpful to predict the occurrenc
出处
《临床检验杂志》
CAS
CSCD
2016年第2期110-113,117,共5页
Chinese Journal of Clinical Laboratory Science
基金
甘肃省技术研究与开发专项(1105TCYA002)
甘肃省卫生行业科研计划管理项目(GWGL2014-02)
