人类mdr1基因新型Taq Man-MGB探针实时定量PCR检测方法的建立

Quantification of Human mdr1 Gene by Using Real Time FQ PCR with Taq Man-MGB Probe

目的建立一种比现有方法敏感、特异性高、重复性好的实时荧光定量PCR检测的新方法检测人类mdr1基因。方法以含有目的基因mdr1cDNA的质粒pHaMDR1/A为阳性模板,用Primerexpress2.0引物设计软件设计引物和TaqMan-MGB探针,建立荧光定量PCR检测方法。结果当待扩增DNA浓度在3.601×103cps/ml～3.601×109cps/ml范围时,模板浓度与循环阈值(Ct)之间的相关性良好,相关系数r2大于0.98。结论应用TaqMan-MGB探针的实时荧光定量PCR方法检测人类mdr1基因,具有灵敏度高、特异性高和精确度高等优点,可作为进一步研究人类mdr1基因的方法。

Objective To establish a novel method for detection of human mdr1 gene. Methods A plasmid pHaMDR1/A with mdrl cDNA was used as a template. Primer Express 2.0 was used for designing of primers and Taq Man-MGB probe for the real-time fluorescent quantitative polymerase chain reaction （FQ-PCR）. Results A good correlation between Ct and the concentration of template was observed when the template concentration was ranged from 3.061×10^5cps/ml to 3.061×10^8 cps/ml （r^2=0.994436）. Conclusion A real-time FQ-PCR based on Taq Man-MGB probe is a sensitive, specific and quantitative method to detect human mdr1.