p53基因第8外显子实时荧光PCR方法的建立

Establishment of real-time PCR for P53 detection

目的建立实时荧光定量聚合酶链反应(PCR)检测p53基因第8外显子的方法。方法用酚-氯仿抽提法提取云南省中医院的常规体检患者的DNA,应用SYBR GreenⅠ作荧光染料,通过检测PCR产物中荧光讯号的强度来定量p53基因,重复测定该标本18次,以此建立检测p53基因的实时荧光定量PCR方法。结果融解曲线分析表明该方法特异可靠。结论该研究建立了p53基因实时荧光定量PCR检测方法,该方法简便、特异、重复性好、可信度高,为p53基因第8外显子的定量研究提供了理想的检测方法。

Objective To establish the real-time PCR for p53 8th extron.Methods DNA was extracted by Trizol from the people having examination in our hospital.SYBR Green Ⅰ was used as staining reagent.p53 8th extron was quantitatively analyzed by determining the intensity of fluorescent signals.The detection was repeated 18 times to establish the needed method.Results Specific and reliable results were achieved.Conclusion A simple,specific,repeatable and reliable method has been established for detection of p53 8th extron.