复合式蝎形引物实时定量检测端粒酶延伸产物  

Real-time Quantitative Assay of Telomerase Product Using the Duplex Scorpion Primer

作　　者：黄艳萍[1] 孔德明[1] 张晓滨[1] 沈含熙[1] 宓怀风[1] 

机构地区：[1]南开大学化学学院高分子功能材料吸附与分离国家重点实验室

出　　处：《化学学报》2004年第3期274-278,共5页Acta Chimica Sinica

基　　金：国家自然科学基金 (No .2 0 0 750 1 2 )资助项目

摘　　要：针对端粒酶延伸产物中靶基因序列的特殊性 ,开发了一种可产生荧光的复合式蝎形引物 ,该引物的 5′端带有可特异性检测靶基因的探针序列 ,PCR阻断剂将其与引物序列连接 .当复合式蝎形引物延伸 ,探针序列与同一分子内的靶基因杂交 ,荧光信号产生 .运用该技术 ,建立了定量检测端粒酶延伸产物的实时荧光PCR方法 .该法可在快速PCR循环条件下 ,对 0 .15～ 1.5 0× 10 3 amol/ μL范围内的样品进行定量检测 ,线性相关系数R2 =0 .9992 .该法操作简便 。To specific target sequence of telomerase product, a fluorogenic duplex scorpion primer has been designed. A probe element attached at the 5′-end of it can specifically detect target gene. A PCR blocker—carbon chain with C 3 group, whose length is as the same as a base pair, is joined between the primer sequence and the probe in the duplex scorpion primer. A fluorescence signal is only produced when the probe sequence is hybridized with the target gene in the extended duplex scorpion primer. Using this technology, a novel method has been developed for quantitative assay of telomerase product by real-time PCR. Accurate quantitative assay can be achieved with sample detected within 0.15～1.50×10 3 amol/μL under fast cycling conditions. The linear correlation factor R 2=0.9992. The method is specific, simple and without post-PCR manipulation.

关 键 词：复合式蝎形引物 实时检测 定量检测 端粒酶 延伸产物 荧光定量PCR技术 恶性肿瘤 诊断 

分 类 号：O657.3[理学—分析化学]