Telomere length detection method and kit

Abstract

The invention provides a human chromosome telomere length rapid quantitative detection method and a kit thereof. The method is characterized in that the chromosomal telomere DNA length and changing rate are accurately measured. The basic principle is that the temperature of the chromosome telomere DNA is quickly changed from 98 DEG C to 50 DEG C, to achieve rapid cooling of the temperature and to achieve the purpose of hybridization of a liquid phase double-stranded probe and nucleic acid molecules. By detecting the fluorescence value before and after the hybridization, the fluorescence change value is calculated and converted to the fluorescence change amount corresponding to the unit DNA to be detected, that is, the change amount of the fluorescence value detected before and after the hybridization respectively is divided by the amount of the DNA for the hybridization, to indicate the relative length of the telomere DNA. According to the method, the hybridization temperature and temperature dropping time are strictly controlled; the renaturation of the telomere DNA is minimized, so that the telomere DNA is more fully hybridized with the probe. Therefore, the method has the advantages of being accurate, fast, cheap, convenient, efficient, time-saving and effort-saving, and providing a simple and practical technical method for the telomere DNA length detection.

Description

technical field [0001] The invention relates to a technical method for rapidly quantitatively detecting the average length of chromosomal telomeric DNA from peripheral blood DNA and a kit thereof, belonging to the field of molecular biology detection. Specifically, it involves a technique for measuring the average length of telomere DNA through competitive hybridization between a simple fluorescent probe and telomere DNA and quantitatively measuring the fluorescent signal of the probe hybridization, which can be applied to the detection of human biological age and human chronic diseases. The assessment and the detection and evaluation of the pharmacodynamics of pro-aging and anti-aging drugs. Background technique [0002] Telomere (Telomere) is a nucleic acid-protein complex composed of a small piece of special telomere DNA and its binding protein that exists at the end of eukaryotic cell chromosomes. It is one of the three major elements that keep chromosomes intact and sta...

AI Technical Summary

Problems solved by technology

The telomere sequence measured by this technique also includes the sequence of the sub-telomeric region, which cannot provide the actual length of the telomere, nor can it accurately quantify the length of the telomere

At the same time, this technique is time-consuming and labor-intensive, and requires a large amount of DNA

[0011] 2. flow-FISH, which combines flow cytometry and fluorescence in situ hybridization, but the technology is complicated, and there are inherent contradictions between cell fixation and complete denaturation of chromosomal DNA, and the number of hybridized probes entering cells is related to the number of unhybridized probes washed out The integrity of the cells cannot be controlled, and the cells need to be kept intact throughout the experimental process for analysis, and the quantification is not accurate enough

Telomere length, although this method has high sensitivity, but because the telomeric DNA sequence is a 6-nucleotide repeat sequence with nearly thousands of copies, the products that appear during the amplification process are extremely complex, and the repeat sequence itself is difficult to effectively amplify, so Quantitative results are unconvincing and lack standardization

[0013] In summary, although telomere DNA length is the most important and accurate indicator of a person's aging speed, there is still a lack of effective quantitative detection technology for telomere DNA length at home and abroad.