摘要
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.
出处
《临床军医杂志》
CAS
2008年第3期442-445,共4页
Clinical Journal of Medical Officers
基金
解放军福州总医院基金资助(200731)