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Telomerase activity detecting method based on fluorescence resonance energy transfer (FRET)

An activity detection, telomerase technology, applied in the direction of microbial assay/inspection, biochemical equipment and methods, etc., to maintain integrity and stability, avoid false positive signals, excellent biocompatibility and nanoscale controllable effect

Active Publication Date: 2019-06-18
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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Problems solved by technology

Therefore, many cannot avoid high false positive signals originating from nuclease digestion, protein binding, or thermodynamic fluctuations

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  • Telomerase activity detecting method based on fluorescence resonance energy transfer (FRET)
  • Telomerase activity detecting method based on fluorescence resonance energy transfer (FRET)
  • Telomerase activity detecting method based on fluorescence resonance energy transfer (FRET)

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Embodiment Construction

[0025] The present invention will be further described in detail below in conjunction with the embodiments, so that those skilled in the art can implement it with reference to the description.

[0026] It should be understood that terms such as "having", "comprising" and "including" used herein do not exclude the presence or addition of one or more other elements or combinations thereof.

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Abstract

The invention discloses a telomerase activity detecting method based on fluorescence resonance energy transfer (FRET). The telomerase activity detecting method comprises the following steps that firstly, fluorescence-modified Flare DNA is designed; then single-stranded DNA S1, single-stranded DNA S2, single-stranded DNA S3 and single-stranded DNA S4 are utilized to construct a DNA tetrahedral structure; then a DNA tetrahedron and the Flare DNA are hybridized, and thus a fluorescent DNA nano-probe is obtained; and the fluorescent DNA nano-probe and telomerase are mixed for reaction, and thus afluorescence spectrum of a mixture is obtained. The designed fluorescent probe based on FRET can provide a precise detection result, false positive signals are avoided, and the influence of system fluctuation in bioimaging is minimized; and on the other hand, a three-dimensional (3D) DNA nanostructure has excellent biocompatibility and nanoscale controllability and can enter cells through a rapidendocytic pathway, the basic integrity and stability of the cells are kept, and the 3D DNA nanostructure can be applied to biosensing and bioimaging after being reasonably assembled.

Description

technical field [0001] The invention relates to the technical field of telomerase detection, in particular to a method for detecting telomerase activity based on FRET. Background technique [0002] Telomerase is considered to be a key biomarker for early cancer diagnosis, and the development of sensitive and practical detection methods for telomerase activity is of great significance for clinical diagnosis, biological research and development of anticancer drugs. The polymerase chain reaction-based telomeric repeat amplification protocol (TRAP) is the most commonly used method for the detection of telomerase, and many other applications, such as fluorescence, colorimetry, electrochemical and electrochemiluminescent techniques, have been developed to Detection of telomerase activity. Although these methods have good sensitivity, most require cell lysis of the sample, resulting in artificial loss of cell origin, and suffer from complicated manipulations and false positive rea...

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Application Information

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IPC IPC(8): C12Q1/6818C12Q1/48
Inventor 缪鹏孟凡渝陈锡峰马筱一梅茜
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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