Seven novel probe systems for real-time PCR provide absolute single-base discrimination, higher signaling, and generic components.
Seven novel probe systems for real-time PCR provide absolute single-base discrimination, higher signaling, and generic components.
复制标题
七种新颖的实时 PCR 探针系统提供绝对的单碱基区分、更高的信号传导和通用组件。
DOI：
10.1016/j.jmoldx.2014.06.008
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发表时间：
2014
期刊：
影响因子：
--
通讯作者：
Shafer,DavidA
中科院分区：
文献类型：
--
作者：
Murray,JamesL;Hu,Peixu;Shafer,DavidA
We have developed novel probe systems for real-time PCR that provide higher specificity, greater sensitivity, and lower cost relative to dual-labeled probes. The seven DNA Detection Switch (DDS)-probe systems reported here employ two interacting polynucleotide components: a fluorescently labeled probe and a quencher antiprobe. High-fidelity detection is achieved with three DDS designs: two internal probes (internal DDS and Flip probes) and a primer probe (ZIPR probe), wherein each probe is combined with a carefully engineered, slightly mismatched, error-checking antiprobe. The antiprobe blocks off-target detection over a wide range of temperatures and facilitates multiplexing. Other designs (Universal probe, Half-Universal probe, and MacMan probe) use generic components that enable low-cost detection. Finally, single-molecule G-Force probes employ guanine-mediated fluorescent quenching by forming a hairpin between adjacent C-rich and G-rich sequences. Examples provided show how these probe technologies discriminate drug-resistantMycobacterium tuberculosismutants,Escherichia coliO157:H7, oncogenicEGFRdeletion mutations, hepatitis B virus, influenza A/B strains, and single-nucleotide polymorphisms in the humanVKORC1gene.
