Method for detecting hotspot mutation of human TERT gene promoter

A technology of TERT-124 and TERT-146, applied in the field of real-time fluorescent quantitative PCR detection, can solve the problems of high detection cost, expensive detection equipment, complicated operation, etc., achieving high universality, clear and objective interpretation of results, and high detection sensitivity. Effect
CN111424094AInactive Publication Date: 2020-07-17AEROSPACE CENT HOSPITAL
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
AEROSPACE CENT HOSPITAL
Publication Date
2020-07-17
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention discloses a method for detecting hot spot mutation of a human TERT gene promoter. The method comprises the following steps: a, enriching templates by adopting DNA enrichment conventionalPCR aiming at different specimens; b, purifying the PCR product, determining the concentration by using an ultraviolet spectrophotometer, and performing concentration calibration and standard substance preparation; and c, carrying out TERT-146 (C / T) and TERT-124 (C / T) mutation rate quantitative detection on the calibrated specimens by adopting qPCR. According to the method, a qPCR technology platform which is conventionally developed clinically is adopted, the high-sensitivity TERT hot spot mutation rate quantitative detection method is established for DNA specimens from various sources, thelower limit of the detected mutation rate reaches 0.05%, and the method has the advantages of being high in detection sensitivity, high in specificity, capable of accurately and quantitatively detecting the mutation rate, low in cost, suitable for conventional clinical development and the like.
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Description

technical field

[0001] The invention relates to the field of biotechnology, in particular to a real-time fluorescence quantitative PCR (qPCR) detection method for detecting the mutation rates of human TERT gene promoter hot spot mutations -146 (C / T) and -124 (C / T). Background technique

[0002] Telomere refers to a unique structure composed of DNA and related proteins containing simple repeating 5'-TTAGGG-3' base series at the end of chromosomes. It exists in eukaryotic cells and its main function is to maintain the integrity of chromosome structure and genome. Stability. With each cell division cycle, telomere DNA is reduced, and telomere length is progressively shortened, eventually leading to the cessation of cell division, forcing cells to senesce and die. Telomerase is a ribonucleoprotein polymerase mainly composed of telomerase RNA template (hTR), telomerase reverse transcriptase (TERT) and telomerase-related proteins. The activation of telomerase is the main mechanis...

Claims

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