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目的建立实时荧光定量PCR法检测mi R-499并探讨其在诊断急性心肌梗死中的应用价值。方法设计mi R-499的颈环结构反转录引物和PCR扩增引物,通过Taq Man探针时荧光定量PCR法检测急性心肌梗死(acute myocardial infarction,AMI)、不稳定心绞痛(unstableanginapectoris,UA)和健康对照各40例血清标本中mi R-499的拷贝量,分析mi R-499的对诊断急性心肌梗死的作用。结果实验的扩增效率为0.94,相关系数为0.992915。AMI组mi R-499的表达水平明显升高,UA组及健康对照组mi R-499的表达水平未见明显升高。结论成功建立了Taq Man探针荧光定量PCR法检测人血清中mi R-499含量的平台,mi R-499有可能成为一种新型且敏感的诊断急性心肌梗死的生物标志物。
Objective To establish a real-time fluorescence quantitative PCR method to detect mi R-499 and explore its value in the diagnosis of acute myocardial infarction. Methods The miR-499 reverse-transcription primers and PCR amplification primers were designed and used to detect acute myocardial infarction (AMI) and unstable anginapectoris (UA) by Taq Man probe. And healthy controls in 40 serum samples of mi R-499 copies of the analysis of mi R-499 on the diagnosis of acute myocardial infarction. Results The experimental efficiency was 0.94 and the correlation coefficient was 0.992915. The expression of mi R-499 in AMI group was significantly higher than that in the UA group and healthy control group. Conclusion The TaqMan probe was successfully established to detect the content of mi R-499 in human serum. Mi R-499 may become a new and sensitive biomarker for the diagnosis of acute myocardial infarction.