Kit for detecting hot spot mutation of human TERT gene promoter

A TERT-124 and promoter technology, applied in the biological field, can solve the problems of complex operation, high detection cost, expensive detection equipment, etc., and achieve the effects of high detection sensitivity, high universality, and clear and objective interpretation of results.

Pending Publication Date: 2020-01-24
AEROSPACE CENT HOSPITAL
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, for the detection of the hotspot mutation rate of TERT, there are NGS method (the lower limit of detection is about 0.1%) and ddPCR method (the lower limit of detection is about 0.1%-0.05%) reported in the open literature. Problems such as high cost and expensive testing equipment are not suitable for clinical routine development

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  • Kit for detecting hot spot mutation of human TERT gene promoter
  • Kit for detecting hot spot mutation of human TERT gene promoter
  • Kit for detecting hot spot mutation of human TERT gene promoter

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Experimental program
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Embodiment Construction

[0023] The present invention will be further described below in conjunction with the accompanying drawings and embodiments.

[0024] The self-prepared instruments required by the kit of the present invention are as follows: various fluorescent quantitative PCR instruments, various conventional PCR thermal cyclers, ultraviolet spectrophotometers, high-speed centrifuges, vortex oscillators, and sterilized water for injection.

[0025] Self-prepared reagents required for the kit of the present invention: DNA extraction kit, common DNA product purification reagent.

[0026] 1. DNA enrichment routine PCR reaction

[0027] (1) Mixed reagents for PCR reaction: various substances at the following concentrations are added to the premixed reagents:

[0028] name Dosage (final concentration) Mastermix 12.5μl(-) 163-F (SEQ ID NO.1) primer 2μl (0.4μM) 163-R (SEQ ID NO.2) primer 2μl (0.4μM) Sterile water for injection 6.5μl(-) DNA template 2μl...

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Abstract

The invention discloses a kit for detecting hot spot mutation of a human TERT gene promoter. The method is characterized in that the kit is characterized by comprising a DNA enrichment conventional PCR premixing reagent for detecting mutation sites of TERT gene promoters-124 (C/T) and-146 (C/T), -124 (C/T) and-146 (C/T) mutant plasmids, a human peripheral blood wild type DNA standard substance, and -124 (C/T) and-146 (C/T) mutation rate detection qPCR premix reagents. The kit provided by the invention adopts a fluorescent quantitative PCR technology platform which is conventionally developed clinically, is used for detecting DNA specimens from various sources, and has the characteristics of high detection sensitivity, strong specificity, accurate quantitative detection of mutation rate, low cost, suitability for conventionally developing clinically and the like.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to a method for detecting human TERT gene promoter hotspot mutations -146(C / T) and -124(C / T) for specimen enrichment, standard product preparation and specimen calibration, and mutation rate detection real-time fluorescent quantitative PCR (qPCR) detection kit. Background technique [0002] Telomere (telomere) refers to the unique structure composed of DNA and related proteins containing a simple repeat 5'-TTAGGG-3' base series at the end of the chromosome. It exists in eukaryotic cells and its main function is to maintain the integrity of the chromosome structure and the integrity of the genome. Stability, every cell division cycle, telomere DNA will decrease, telomere length shortens progressively, eventually leading to the cessation of cell division, forcing cells to age and die. Telomerase is a ribonucleoprotein polymerase mainly composed of telomerase RNA template (hTR), t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851
CPCC12Q1/6886C12Q1/6851C12Q2600/156C12Q2600/118C12Q2563/107C12Q2561/113C12Q2531/113
Inventor 梁国威张洁
Owner AEROSPACE CENT HOSPITAL
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