Exciton Primer-Mediated SNP Detection in SmartAmp2 Reactions
Exciton Primer-Mediated SNP Detection in SmartAmp2 Reactions
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DOI：
10.1002/humu.21177
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发表时间：
2010-02-01
期刊：
影响因子：
3.9
通讯作者：
Hayashizaki, Yoshihide
中科院分区：
文献类型：
--
作者：
Lezhava, Alexander;Ishidao, Takefumi;Hayashizaki, Yoshihide
DNA检测中最常用的嵌入荧光染料缺乏任何序列特异性，而所谓的激子引物可以通过充当“序列特异性染料”来克服这种限制。“与互补序列杂交后，激子引物的荧光提供了序列特异性信号，用于实时监测扩增反应。应用于Smart突变检测过程，激子引物显示出高信号强度和低背景，与SYBR绿色I相比，具有上级特异性和灵敏度。在一个激子引物中使用多种染料或在同一扩增反应中使用多种激子引物可以进一步增强信号强度。在这里，我们展示了使用激子引物对VKORC 1基因座的单核苷酸多态性（SNP）进行基因分型。（-1639 G>A）与Warpon给药相关，作为SmartAmp 2的激子引物介导的基因分型的实例基因分型测定可仅使用一种标记的激子引物进行终点检测，或同时通过真实的，时间监测使用具有不同染料的两种激子引物在单管反应中检测野生型和突变体等位基因。SmartAmp 2的激子打印机介导的基因分型直接从血液样本工作，为快速床旁检测提供了上级解决方案。Mutat 31：208-217，2010. (C)2010 Wiley-Liss，Inc.
Most commonly used intercalating fluorescent dyes in DNA detection are lacking any sequence specificity, whereas so,called Exciton Primers can overcome this limitation by functioning as "sequence-specific dyes." After hybridization to complementary sequences, the fluorescence of Exciton Primers provides sequence-specific signals for real-time monitoring of amplification reactions. Applied to the Smart mutation detection process, Exciton Primers show high signal strength with low background leading to a superior specificity and sensitivity compared to SYBR Green I. Signal strength can be further enhanced using multiple dyes within one Exciton Primer or use of multiple Exciton Primers in the same amplification reaction. Here we demonstrate the use of Exciton Primers for genotyping a single nucleotide polymorphism (SNP) in the VKORC1 locus (-1639G>A) relevant for Warfarin dosing as an example for Exciton Primers mediated genotyping by SmartAmp2 The genotyping assay can use only one labeled Exciton Primer for endpoint detection, or simultaneously by real,time monitoring detect wild-type and mutant alleles in a one-tube reaction using two Exciton Primers having different dyes. Working directly from blood samples, Exciton Printer mediated genotyping by SmartAmp2 offers superior solutions for rapid point-of-care testing. Hum Mutat 31:208-217, 2010. (C) 2010 Wiley-Liss, Inc.