Method for immediate detection of telomerase activity based on gas pressure change caused by H2O2 (hydrogen peroxide) decomposition catalyzed by platinum nanoparticle

A platinum nanoparticle and air pressure change technology, which is applied in biochemical equipment and methods, microbial determination/inspection, and measurement devices, can solve the problems of high cost, poor practicability, and complicated operation of telomerase activity detection

Active Publication Date: 2017-06-06
SHAANXI NORMAL UNIV
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  • Abstract
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Problems solved by technology

[0004] In order to overcome the defects of high cost, complicated operation and poor practicability in the prior art of telomerase activity detection, the present invention provides a catalyst based on platinum nanoparticles, which can be instant and sensitive without polymerase chain reaction. Method for detecting telomerase activity

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  • Method for immediate detection of telomerase activity based on gas pressure change caused by H2O2 (hydrogen peroxide) decomposition catalyzed by platinum nanoparticle
  • Method for immediate detection of telomerase activity based on gas pressure change caused by H2O2 (hydrogen peroxide) decomposition catalyzed by platinum nanoparticle

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Embodiment 1

[0017] Taking the detection of telomerase in HeLa cells as an example, the specific detection method is as follows:

[0018] 1. Take 0.48 μL of aminated magnetic beads with a concentration of 50 μg / μL (NH2-MB, manufactured by Shaanxi North American Gene Co., Ltd.), wash twice with phosphate buffer solution, and then add the cleaned aminated magnetic beads to 120 μL of phosphate buffer solution, and add 1.2 μL of 10 μmol / L formylated telomerase primer (provided by Sangon Bioengineering (Shanghai) Co., Ltd., wherein the sequence of the telomerase primer is: 5'-AAT CCG TCG AGC AGA GTT-3'), shaking at 30°C for 2 hours, and then washing with phosphate buffer solution by magnetic separation to remove telomerase primers not connected to the surface of magnetic beads to obtain magnetic beads / telomerase Primer complex: Dilute the magnetic bead / telomerase primer complex to 120 μL with phosphate buffer solution, and store it in the refrigerator at 4°C for later use.

[0019] 2. After mi...

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Abstract

The invention discloses a method for immediate detection of telomerase activity based on gas pressure change caused by H2O2 (hydrogen peroxide) decomposition catalyzed by a platinum nanoparticle. The method comprises the steps: when active telomerase exists, a telomerase primer modified on the surface of a magnetic bead is bonded to telomerase, thus the telomerase primer is extended along a 3' end of the telomerase primer and a DNA single strand with TTAGGG repeats is formed; the long DNA single strand can hybridize with multiple cDNAs, and by modifying the surface of the platinum nanoparticle with the cDNA, the platinum nanoparticle is fixed to the surface of the magnetic bead; then the platinum nanoparticle is utilized to catalyze H2O2 decomposition to produce oxygen, and the gas pressure of a system changes; then a portable barometer is adopted for detecting a gas pressure value of the system, so that the immediate detection of the telomerase activity can be realized. The detection method disclosed by the invention is simple, rapid and sensitive, and can be used for detecting the expression level of telomerase activity in different tumor cells, and the detection method is of great significance in the diagnosis and treatment of cancers with telomerase as a target point.

Description

technical field [0001] The invention belongs to the technical field of tumor activity detection, and in particular relates to a simple and sensitive method for real-time detection of telomerase activity in cancer cells based on changes in air pressure. Background technique [0002] Telomere is a special structure located at the end of chromosomes in eukaryotic cells. It is composed of telomere DNA and telomere-related proteins. The telomere DNA is composed of non-coding repeat sequences. Different species have different repeat sequences, such as human telomere The DNA is composed of 5'-TTAGGG-3' repeat sequence, and the length ranges from 4 to 15kb. The main function of telomeres is to protect the ends of chromosomes, prevent chromosome fusion, recombination and degradation. In normal cells, as the cells proliferate, the telomeres will gradually shorten. When the telomeres shorten to a certain extent, the chromosomes will become unstable, and the cells will age and die. Te...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/48
CPCC12Q1/48G01N2333/9128
Inventor 金燕王彦君李保新杨璐竹
Owner SHAANXI NORMAL UNIV
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