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首页> 外文期刊>Journal of Pharmaceutical and Biomedical Analysis: An International Journal on All Drug-Related Topics in Pharmaceutical, Biomedical and Clinical Analysis >Typing and determination of SNP functional gene based on highly selective and signal-amplified fluorescence double-probe with the help of ExoIII nuclease and magnetic bead
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Typing and determination of SNP functional gene based on highly selective and signal-amplified fluorescence double-probe with the help of ExoIII nuclease and magnetic bead

机译：基于高选择性和信号扩增的荧光双探针的SNP官能基因的键入和测定尤其是exoIII核酸酶和磁珠

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摘要

We have developed a fluorescence double-probe detection system with signal amplification for simple typing and determination of single nucleotide polymorphism (SNP) functional gene based on non-sequence dependence of ExoIII nuclease on dsDNA and rapid separation of magnetic bead. Matched detected gene can cyclically release abundant fluorescence-labeled ssDNA from the probe and the corresponding measured fluorescence signal is amplified up to 6063 times. In this case, the probe cannot release the measured fluorescence signal for the point mutation gene and then the corresponding measured signal is inhibited. According to signal amplification and inhabitation of the probe, we proposed both an accurate genotyping approach with strong specificity and a sensitive determination approach with high selectivity for SNP functional gene. For qualitative genotyping, there are obvious genotype-based differences of measured fluorescence phenotypes among three kinds of the samples of the investigated SNP. The quantitative determinations of its wild-type gene and mutant gene have all a good linearity in the range from 0.5 to 500 pmol/L with the correlation coefficients R-2 of 0.9940 and 0.9911, and a high sensitivity with the detection limits of 0.11 and 0.20 pmol/L, respectively. Compared to the usual single-probe detection system, the developed double-probe system can achieve not only accurate genotyping but also the sensitive gene determination. Meanwhile, it is also a simple and reliable method for both quantitative and qualitative analysis of functional gene. (C) 2019 Elsevier B.V. All rights reserved.
机译：我们开发了一种荧光双探针检测系​​统，具有信号放大，用于基于EXOIII核酸酶对DSDNA对exoIII核酸酶的非序列依赖性的单核苷酸多态性（SNP）功能基因的简单键入和测定。匹配的检测到的基因可以从探针循环循环释放丰富的荧光标记的SSDNA，并且相应的测量荧光信号被扩增至6063倍。在这种情况下，探针不能释放点突变基因的测量荧光信号，然后抑制相应的测量信号。根据信号放大和探针的鉴别，我们提出了一种具有强大特异性的准确基因分型方法和具有高选择性的SNP官能基因的敏感测定方法。对于定性基因分型，存在明显的基于基因型的基于基因型的测量荧光表型在研究的S​​NP样品中的三种样品中的差异。其野生型基因和突变基因的定量测定在0.5至500pmol / L的范围内具有0.9940和0.9911的相关系数R-2的良好线性度，以及高灵敏度，检测限为0.11和0.20 pmol / l。与通常的单探针检测系​​统相比，开发的双探针系统不仅可以达到准确的基因分型，而且达到敏感的基因测定。同时，它也是一种简单可靠的功能基因定量和定性分析的方法。 （c）2019 Elsevier B.v.保留所有权利。

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