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TP53 detection method based on exonuclease III and ribozyme double signal amplification strategy

An exonuclease and dual signal technology, which is applied in the field of TP53 detection based on the exonuclease III and ribozyme dual signal amplification strategy, can solve the problems of high instrument requirements, low sensitivity and high cost, and achieve high sensitivity. , strong specificity, low cost effect

Inactive Publication Date: 2019-02-26
SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI
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Problems solved by technology

Traditional TP 53 detection methods include high performance liquid chromatography, denaturing gradient gel electrophoresis, and immunohistochemistry. However, these methods face many problems, such as high cost, high instrument requirements, low sensitivity, and complicated operation.

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  • TP53 detection method based on exonuclease III and ribozyme double signal amplification strategy
  • TP53 detection method based on exonuclease III and ribozyme double signal amplification strategy
  • TP53 detection method based on exonuclease III and ribozyme double signal amplification strategy

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Embodiment Construction

[0024] 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.

[0025] 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.

[0026] like figure 1 , a kind of TP 53 detection method based on exonuclease III and ribozyme double signal amplification strategy of the present embodiment, this method first designs the DNA probe of neck ring DNA (DNA probe a) and fluorescent dye label; Then DNAprobe b is modified to the surface of gold nanoparticles, and the fluorescence of the fluorescent dye is quenched due to its proximity to the gold nanoparticles; when the gene TP 53 to be detected exists, TP 53 can be complementary paired with DNA probe a, forming a blunt end at the 3' end. At this time, exonuclease III will cut DN...

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Abstract

A TP53 detection method based on an exonuclease III and ribozyme double signal amplification strategy is disclosed. The method includes 1) preparing gold colloid containing gold nanoparticles; 2) modifying the gold nanoparticles with a DNA probe; 3) mixing DNA with a neck ring structure and TP 53 into a buffer solution, and adding exonuclease III to prepare a reaction solution; 4) mixing the reaction solution with the gold colloid solution modified with the DNA probe, and adding zinc chloride for further reaction; and 5) measuring fluorescence signals. Through the exonuclease III and ribozymedouble signal amplification strategy, the detection limit of the TP 53 that is a substance to be detected is greatly reduced. The method is highly sensitive, rapid and cheap when being used for TP 53detection, the detection limit is 21 fM, and TP 53 biosensing application can be achieved by combining the exonuclease III and ribozyme double signal amplification strategy and a fluorescence spectrumtechnique.

Description

technical field [0001] The invention relates to the field of nucleic acid detection, in particular to a TP 53 detection method based on the double signal amplification strategy of exonuclease III and ribozyme. Background technique [0002] Human TP 53 is located on chromosome 17, and the TP 53 gene in cancer patients often undergoes significant mutations. Studies have shown that TP 53 plays an important role in the occurrence and progression of breast cancer and ovarian cancer. TP 53 can not only be used as a marker for early diagnosis of cancer, but also can be used to monitor the development of cancer and the prognosis of cancer patients. Therefore, the development of a rapid and accurate detection method for TP 53 has significant significance in both biology and clinical diagnosis. Traditional TP 53 detection methods include high performance liquid chromatography, denaturing gradient gel electrophoresis, and immunohistochemistry. However, these methods face many problem...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/682
CPCC12Q1/682C12Q2521/319C12Q2563/107C12Q2521/337C12Q2563/137
Inventor 杨大威缪鹏陈锡峰孟凡渝
Owner SUZHOU INST OF BIOMEDICAL ENG & TECH CHINESE ACADEMY OF SCI