荧光实时定量PCR检测老年人外周血淋巴细胞β_1和β_2肾上腺素能受体基因mRNA表达

Fluorescent Real-time Quantitative PCR for Detection of β_1-and β_2-AR mRNA Expressions in Elderly Patients'Peripheral Lymphocytes

目的建立荧光实时定量PCR法检测老年人外周血淋巴细胞β1和β2肾上腺素能受体（AR）基因mRNA表达。方法随机选择ASAⅠ-Ⅱ级老年患者30例，分离外周静脉血淋巴细胞，采用荧光实时定量PCR法检测β1和β2-AR基因mRNA表达，并与半定量PCR检测结果进行比较。结果30例患者外周血淋巴细胞荧光实时定量PCR均检出β1和β2-ARmRNA表达，不同性别组同-基因表达量无显著差异（P〉0.05）；荧光实时定量PCR检测β2-AR mRNA表达量显著高于β1—AR mRNA（P〈0．001），而半定量PCR随着循环次数增加，β1和β2-ARmRNA表达差异减小。结论所建立的荧光实时定量PCR成功检测老年人外周血淋巴细胞β1和β2-AR基因mRNA表达，有较高的灵敏性及特异性；半定量PCR扩增平台期影响试验结果判断。

Objective To establish a real-time quantatitive polymerase chain reaction （PCR） method for the detection of β1-and β2-AR mRNA expression in elderly patients＇ peripheral lymphocytes. Methods Thirty elderly patients with ASA grade Ⅰ - Ⅱ were recruited. Their blood was collected and the lymphoctes were isolated. The expressions of β1-and β2-AR mRNA were detected with fluorescent quantitative real-time PCR and semi-quantitive PCR. Results There were expressions of β1- and β2-AR mRNA in all the subjects with real-time PCR, and the latter was significally more abundant than the former （ P 〈 0. 001 ）. Also, there was no difference between different genders （ P 〉 0.05 ）. With semi-quantive PCR, the ratio of β2-AR mRNA/β1-AR mRNA decreased when the cycles increased （ P 〈 0.05）. Conclusion Semi-quantatitive PCR is not perfect at the platform stage, but fluorescent real-time quantitative PCR is more accurate and sensitive than semiquantitive PCR for the detection of β1- and β2-AR mRNA expressions in elderly patients＇ peripheral lymphocytes.