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Method and system for acquiring individual age of Chinese population and amplification detection system

An individual and population-based technology, applied in biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., can solve problems such as need for improvement, questionable stability, and inability to explain age variation well.

Active Publication Date: 2019-09-20
HUAZHONG UNIV OF SCI & TECH
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The study found that although mitochondrial DNA 4977bp deletion, advanced glycation end products, aspartate racemization, and telomeric restriction fragment length all showed significant age-dependent changes, these markers were inferred with low precision , poor accuracy and repeatability of measurement results
In addition, messenger RNA is less biologically and chemically stable than DNA molecules, so it is less used in age inference studies
In 2010, Zubakov et al first developed a more reliable age estimation method for blood samples by using signal-binding T-cell receptor deletion ring markers, but its stability is still questioned, and it is not suitable for other types of samples
In 2014, Florath et al. also constructed an age inference model containing 17 AR-CpGs, and the MAD was 2.6 years old. However, compared with the previous two studies, these sites could not explain the age variation well (R 2 =0.71)
[0009] In summary, a high-precision age estimation model that can be applied to both male and female individuals in the Chinese population has not yet been established, and the accuracy of the existing age estimation models applicable to female individuals still needs to be improved, and other CpG sites can also be used. Points to build a high-precision age inference model

Method used

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  • Method and system for acquiring individual age of Chinese population and amplification detection system
  • Method and system for acquiring individual age of Chinese population and amplification detection system
  • Method and system for acquiring individual age of Chinese population and amplification detection system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0100] Example 1 Screening of AR-CpGs

[0101] 1 sample

[0102] 42 unrelated healthy volunteers of Chinese Han nationality, including 14 young people (Group A, 18-22 years old), middle-aged group (B group, 36-43 years old) and elderly group (C group, 58-62 years old) each, And there are 7 males and 7 females in each group. 2-5 ml of peripheral blood was collected by venipuncture, anticoagulated with EDTA, and stored in a -70°C ultra-low temperature refrigerator for later use.

[0103] 2 Methylation Detection

[0104] The methylation detection is entrusted to Jingneng Biotechnology (Shanghai) Co., Ltd., which includes the following steps:

[0105] (1) DNA extraction and quantification: 200 μl of peripheral blood genomic DNA was extracted using QIAamp DNA Blood Mini Kit (QIAGEN, Germany), and eluted with 70 μl of Buffer AE. Using Quant-iT TM PicoGreen TM dsDNA AssayKit (Thermo Fisher Scientific, USA) was used for DNA quantification. 50 ng of genomic DNA was checked by ...

Embodiment 2

[0116] Verification of Example 2 Candidate AR-CpGs

[0117] 1 sample

[0118] 60 unrelated healthy individuals of Chinese Han nationality, including 20 young people (18-23 years old), 20 middle-aged people (38-43 years old) and 20 old people (55-61 years old) in each group, and 10 males and 10 females in each group. Actual age is equal to the number of days from the sample collection date to the date of birth recorded on the ID card, birth certificate or household registration book divided by 365, and 2 decimal places are retained. 2-5 ml of peripheral blood was collected by venipuncture, anticoagulated with EDTA, and stored in a 4°C refrigerator for later use.

[0119] 2 Methylation detection

[0120] Male samples were only used to detect male candidate AR-CpGs, and female samples were only used to detect female candidate AR-CpGs.

[0121] (1) DNA extraction and quantification: Genomic DNA in 200 μl of peripheral whole blood of the individual was extracted using QIAamp DNA...

Embodiment 3

[0140] Example 3 Selection of AR-CpGs in Candidate Regions and Establishment of Multiple Linear Regression Model

[0141] 1 sample

[0142] 308 unrelated healthy individuals of Chinese Han nationality (1-85 years old), including 141 females (3-80 years old) and 167 males (1-85 years old). The same method as in Example 2 was used for age calculation and peripheral blood collection.

[0143] 2 Methylation detection

[0144] Male samples are only used to detect male candidate regions, and female samples are only used to detect female candidate regions. The same method as in Example 2 was used to complete the methylation detection and data extraction of CpG sites in the candidate region. Since cg22454769 (FHL2) has been reported by multiple studies as a site that is not affected by gender, the female candidate region F2 is also used as one of the male candidate regions. Here, 500-1000 ng of genomic DNA was transformed using EpiTect Fast DNA Bisulfite Kit, and eluted with 15 μl...

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Abstract

The invention belongs to the technical field of biology and relates to a method and system for acquiring the individual age of Chinese population and an amplification detection system. The method includes: extracting the genome DNA of an individual; performing hydrosulphite treatment on the DNA; acquiring the methylation rate of one group of CpG sites in the DNA; using R software to perform regression analysis on the methylation rate of the CpG sites and the age of the individual to build a regression model for inferring the age of the individual. The method and system has the advantages that the individual age of the Chinese population can be accurately inferred, and mean absolute deviation of the inferred age is 2-4; the method and system is especially applicable to police practice, age inferring is performed through the individual blood or blood stain sample extracted in a crime scene, the age range of a suspect or a victim can be obtained, information can be provided for case investigation, the search range can be narrowed, and case cracking speed can be increased.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method, a system and an amplification detection system for obtaining the individual age of a Chinese population. Background technique [0002] Age is an individual characteristic with a biological basis. When the fingerprint database, reference sample and DNA database are unable to provide matching information, by accurately inferring the age of the source of the biological sample at the crime scene, the search scope of unknown suspects can be narrowed, the identity of victims of disaster accidents can be assisted, and the identification of age can be improved. Prediction of dependencies on externally visible features such as hair color, baldness, and skin wrinkles. Therefore, the estimation of individual age has always been one of the important research topics in forensic science. Currently, forensic technicians can infer an individual's age by measuring morphologica...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6876C12Q1/6858C12N15/11
CPCC12Q1/6858C12Q1/6876C12Q2600/154C12Q2523/125C12Q2535/122C12Q2565/301C12Q2537/165
Inventor 黄代新肖超易少华黄钰洁陈晓英
Owner HUAZHONG UNIV OF SCI & TECH
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