Nanometer probe for testing DNA through combination of fluorescence intensity and fluorescence polarization

Abstract

The invention discloses a nanometer probe for testing DNA through combination of fluorescence intensity and fluorescence polarization, and belongs to the technical field of nucleic acid detection. The nanometer probe comprises fluorescent protein, a capturing probe, a reporting probe, and nanometer gold particles. The fluorescent protein is taken as a donor and connected with the capturing probe; the nanometer gold particles are taken as a receptor and connected with the reporting probe; the capturing probe and the reporting probe form an energy transfer donor and receptor pair through complementary pairing of parts of basic groups; and the capturing probe and target DNA are in complete complementary pairing. The nanometer probe is combined with fluorescence intensity parameters and fluorescence polarization parameters with high sensitivity, thereby achieving non-amplification DNA high-sensitivity detection; and through utilization of opposite variation tendencies of fluorescence intensity and fluorescence polarization in a process of forming and vanishing resonance fluorescence energy transfer, a specific value of fluorescence intensity to fluorescence polarization is introduced, and a detection signal of DNA is amplified.

Description

technical field [0001] The invention belongs to the technical field of nucleic acid detection, and in particular relates to a nanoprobe for detecting DNA by combining fluorescence intensity and fluorescence polarization. Background technique [0002] Genes are the material basis of hereditary variation and dominate the basic structure and performance of life. Existing gene analysis is aimed at population cells, and the result is only the average value of the cell population signal or the information of the dominant number of cells. However, cells are heterogeneous. Even single cells from the same source are different from each other in many aspects due to random biological processes and environmental disturbances. Compared with the existing gene sequencing technology, single-cell sequencing technology not only helps to understand cell heterogeneity, but also can detect trace gene expressors or rare non-coding RNA, and can also be used to find new cell types, cell subtype i...

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Problems solved by technology

[0003] In 2005, Zhang et al. from Johns Hopkins University in the United States developed a sandwich-type luminescent probe based on quantum dot fluorescent dye and Cy5 dye, which can be used to determine the content of DNA. Unfortunately, this detection requires complex nuclei. Extraction and other pretreatment processes, and have higher requirements for fluorescence photon detection instruments

[0005] In addition to the traditional fluorescence intensity detection index, fluorescence polarization is an intrinsic detection parameter of fluorescent molecules, and there is no report on the detection method combining fluorescence intensity and fluorescence polarization.

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