【摘要】 目的建立具有良好敏感性、重复性的聚丙烯酰胺梯度凝胶法(PAGE),以测定低密度脂蛋白(LDL)颗粒的大小。方法灌注3%～16%的聚丙烯酰胺梯度凝胶,分离LDL颗粒,并与经典的密度梯度超速离心法(DUC)进行比较。对于凝胶法进行板内和板间重复性的测定。随后分别观察-30℃和-70℃下保存24周的血清对采用凝胶法分离LDL颗粒的影响。结果与DUC法相比,PAGE法的敏感度为70%,特异度为100%。凝胶分离LDL亚组分的平均板内差异为0.44%,平均板间差异为2.05%。-30℃和-70℃分别保存2、4、8、16和24周对血清LDL颗粒直径无明显影响。结论本研究建立的3%～16%的PAGE法具有较好的敏感性和重复性,具有临床和科研使用的可行性。更多还原

【Abstract】 Objective To establish a polyacrylamide gel electrophoresis(PAGE)-based method with high sensitivity and repeatability for measuring the size of low-density lipoprotein(LDL)particles.Methods LDL parti- cles were separated using 3%-16%polyacrylamide gradient gel electrophoresis,and the results were compared with those by density gradient ultraeentrifugation(DUC),which was considered as the golden standard.The intra- and inter-gel repeatability of the PAGE-based method was assessed,and the effect of serum preservation at-30℃and-70℃for up to 24 weeks on the size of the separated LDL particles was also evaluated.Results Compared to DUC method,the sensitivity and specificity of gradient PAGE was 70%and 100%,respectively;with the intra- and inter-gel difference being 0.44%and 2.05%,respectively.Preservation of the serum samples at-30℃or -70℃for 2,4,8,16 or 24 weeks did not result in significant variation in the size of the separated LDL particles. Conclusion 3%-16%gradient PAGE established in this study has good sensitivity and specificity for separating the LDL particles and is valuable for clinical and laboratory detection of small dense low-density lipoprotein.更多还原