实时荧光定量PCR在检测人ANP基因表达的应用

Application of Fluorescent Real-Time PCR for the Detection of Expression of Human Atrial Natriuretic Peptide Gene

目的建立心房钠尿肽(ANP)基因mRNA表达水平的实时荧光定量PCR检测方法,并对该方法进行初步评价。方法以基因表达产物为模板,建立实时荧光定量PCR检测方法,对样本中的心房钠尿肽含量进行相对定量,比较不同样本组的基因表达水平。结果所建立的实时荧光定量PCR方法熔解曲线中熔解峰单一。肺癌患者胸腔样本的ANP含量为对照样本的4.68倍,血清样本为对照样本的16.03倍。结论所建立的ANP实时荧光定量PCR检测方法具有较高的特异性。肺癌患者胸腔液和血清中ANP的含量明显增高。

Objective To determine the expression level of human atrial natriuretic peptide （hANP） using fluorescent real-time PCR （RT-PCR）. Methods Based on the GeneBank sequence analysis of hANP gene,the conserved sequence was selected in the design of primers for RT-PCR. The sensitivity was evaluated by the dilution method. Clinical samples of pleural effusion and serum were then analyzed. Results Single melting peak was observed in the RT-PCR melting curve. The expression level of hANP in pleural effusion and serum samples from patients with lung cancer was 4.68-fold and 16.03-fold higher than the control samples,respectively. Conclusion RT-PCR is specific for the detection of hANP expression. The expression level of hANP was high in the patients with lung cancer.