胃肠道恶性肿瘤C-myc基因荧光PCR方法的研究  

Research of fluorogenic quantitative polymerase chain reaction method to quantify C-myc in Gastrointestinal tumor

作　　者：毛海红[1] 徐冉行[1] 刘东海 

机构地区：[1]浙江省宁波市第六医院检验科,315010 [2]浙江省宁波市第五医院,315010

出　　处：《中国优生与遗传杂志》2010年第6期23-26,共4页Chinese Journal of Birth Health & Heredity

摘　　要：目的建立胃肠癌中C-myc基因的荧光定量PCR(FQ-PCR)方法。方法胃肠癌细胞中的C-myc基因与质粒PGEM-T Easy Vector重组,转化大肠杆菌E.coli DH5α,获得克隆的C-myc基因标准模板。用ABI PRISM 7700PCR仪检测FQ-PCR扩增产物制成标准曲线来检测未知标本中C-myc的含量。结果 FQ-PCR扩增产物呈"S"形动力学曲线;ct(循环阈值)与PCR体系中起始模板拷贝数的对数值之间存在严格的线性关系,显示了FQ-PCR定量的准确性。结论 FQ-PCR是一种快速、简便、灵敏、准确的定量C-myc基因的方法。Objective ： To establish a fluorogenic quantitative polymerase chain reaction （ FQ - PCR） method for the routine quantification of C - myc gene in gastrointestinal tumor. Methods： The C - myc standard gene was obtained by in vitro amplification of cloned C - myc fragment in plasmid PGEM - T Easy Vector. FQ - PCR product was detected using a 7700 ABI PRISM sequence detector system and obtained C - myc standard curve to quantity C - myc of unknown sample. Results ： FQ - PCR generated ＂S＂ kinetics curve of PCR amplification constructed by relating the fluorescence signal intensity （ A Rn） to the cycle number; The standard curve of C - myc was constructed by the linear relationship between the cycle threshold （ct） and the log of starting copy number. The high correlation （0. 999） reveal the reliability of FQ - PCR. Conclusion ： The FQ - PCR is a rapid, sensitive, reliable method for quantification of C - myc gene.

关 键 词：胃肠癌 C—myc 荧光定量-PCR 

分 类 号：R735[医药卫生—肿瘤]