Telomere length detection kit and method and biological age evaluation method thereof

A technology of telomere length and kit, which is applied in the field of telomere length detection kit and biological age evaluation, can solve the difficult problems of continuous dynamic monitoring and accurate comparative analysis of telomere length, and batch-to-batch differences in test results. Achieve the effects of accurate detection, dynamic evaluation and analysis, stable effect, and easy access to raw materials

Pending Publication Date: 2019-11-05
陕西九州医学检验有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In the experiment, due to different personnel or using different batches of reagents for operation, there will be batch-to-batch dif...

Method used

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  • Telomere length detection kit and method and biological age evaluation method thereof
  • Telomere length detection kit and method and biological age evaluation method thereof
  • Telomere length detection kit and method and biological age evaluation method thereof

Examples

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Embodiment 1

[0055] A case of human leukocyte telomere length detection and biological age calculation using the kit of the present invention:

[0056] (1) Use a spectrophotometer to detect the concentration of whole blood genomic DNA after extraction, the concentration of Sample 1 is 51.14ng / μL; dilute the sample DNA concentration to 10ng / μL;

[0057] (2) Configure qPCR reaction solution, sample DNA and 5 quality control products to carry out internal reference qPCR reaction and telomere qPCR reaction respectively, and each reaction is repeated 3 times;

[0058] Wherein, the qPCR reaction system of the internal reference gene comprises: 2×SYBR Green I qPCR Mi×10 μL; 0.8 μL of the forward primer of the internal reference gene; 0.8 μL of the reverse primer of the internal reference gene; wherein, the forward primer of the internal reference gene and the internal reference gene The concentration of the reverse primer was 10 μmol / L; 50×ROX Reference DyeII 0.4 μL; H 2 O 6 μL; Genomic DNA 2 μL...

Embodiment 2

[0071] Using the kit of the present invention to verify the repeatability of the telomere length experiment.

[0072] The T / S ratio calibration calculation results of two samples in 7 independent experiments were compared to verify the repeatability of the experiment. The results showed that the T / S ratio calibrated by quality control had good repeatability in multiple independent experiments. It can effectively reduce the difference between experimental batches, the test result is accurate and reliable, and can be used for continuous monitoring and evaluation of telomere length.

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Abstract

The invention discloses a kit for telomere length detection. The kit includes a telomere gene primer sequence and a reference gene primer sequence. The invention also discloses a biological age evaluation method, which comprises the following steps: (1) establishing a Chinese population telomere length detection calibration T/S database; (2) fitting algorithm according to the above telomere lengthcalibration T/S value-age model distribution diagram, and establishing a telomere length-biological age algorithm model system. With the application of the kit and by using the calibration T/S ratioas the telomere length evaluation standard, accurate detection and dynamic evaluation analysis of telomere length can be realized. With the application of the kit, scientific evaluation of biologicalage can be realized through a biological age database and a calculation model system. With the application of the kit for telomere length detection, the effect is stable, raw materials are cheap and easily-available, and experiment operation is simple and feasible.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a kit for detecting telomere length and a biological age evaluation method. Background technique [0002] Telomeres exist at the ends of chromosomes in eukaryotic cells and are a special structure composed of specific binding proteins and highly repetitive non-transcribed DNA sequences. Telomere DNA consists of two strands, one is the C strand in the 3'-5' direction (AATCCC), and the other is the G strand in the 5'-3' direction (TTAGGG). In addition to the complementary part of the C-G chain, the end of the G chain is 50-300nt longer than the C chain, and the part that grows out of the tail will be folded back and inserted into the complementary sequence of the telomere to combine with the C chain, replacing part of the G chain, forming a D-loop and T- loop structure. The structure of telomeres is stable, just like the caps of chromosomes exist at both ends of chromosomes, protecti...

Claims

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Application Information

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IPC IPC(8): C12Q1/6851
CPCC12Q1/6851
Inventor 张颖聂苏秦俞瑞卿高思欣
Owner 陕西九州医学检验有限公司
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