Composite amplification system for jointly detecting InDel genetic markers of human full locus group, and kit and application thereof

A composite amplification system and a genetic marker technology, which is applied to a composite amplification system and a kit and application field for the combined detection of InDel genetic markers of the human whole locus group, which can solve the problems such as the limitation of forensic medicine efficiency.

Pending Publication Date: 2019-10-15
ACADEMY OF FORENSIC SCIENCE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In addition, the research of forensic DNA laboratories at home and abroad mostly focuses on about 30 Inde

Method used

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  • Composite amplification system for jointly detecting InDel genetic markers of human full locus group, and kit and application thereof
  • Composite amplification system for jointly detecting InDel genetic markers of human full locus group, and kit and application thereof
  • Composite amplification system for jointly detecting InDel genetic markers of human full locus group, and kit and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] This embodiment is a kit containing a multiplex amplification system for joint detection of InDel genetic markers of the human whole locus group and its corresponding detection method. It includes:

[0032] 1. InDel genetic marker screening for forensic applications;

[0033]Based on different databases, screen the InDel genetic markers that are suitable for the main population in China, have high personal identification ability, and cover human autosomes and sex chromosomes. The specific screening principles are as follows: the Human Genome Browser (Genome Browser) in the Galaxy System (Galaxy) developed and maintained by the University of California Santa Cruz (UCSC) Human Genome Bioinformatics Group Browser) as a screening tool, combined with the dbSNP database, screened InDel markers according to the following criteria: ①InDel allele length (inserted or deleted bases) ≥ 1bp and 0.1; ④ scattered in different human autosomes and sex chromosomes; ⑤ the distance betwe...

Embodiment 2

[0052] This example is the application of the kit constructed in Example 1 in the detection of degraded samples.

[0053] 1. Collect formalin-fixed and paraffin-embedded human tissue samples, and extract sample DNA;

[0054] 2. The DNA in step 1 is amplified and detected by using the multiple amplification kit containing 45 InDel markers and the SiFaSTRTM 23-plex kit containing 21 STR loci in Example 1 respectively, and the typing patterns are as follows: figure 2 with image 3 shown.

[0055] A variety of biological specimens in forensic practice are affected by complex factors such as temperature, humidity, biology or chemistry in the environment, and their DNA molecules are easily damaged and DNA strands are broken. Such as figure 2 As shown, the amplification product of the InDel composite amplification kit described in this embodiment is short, which is conducive to the typing detection of degraded samples and provides more DNA genetic evidence; and image 3 The SiF...

Embodiment 3

[0057] This embodiment is the forensic verification work of the kit constructed in embodiment 1.

[0058] The sensitivity, specificity, accuracy, applicability and forensic parameters of the kit constructed in Example 1 were calculated according to the requirements of the Scientific Working Group for DNA Analysis Methods (SWGDAM). The results show that the kit constructed in Example 1 has high sensitivity, and the complete genotyping of 45 InDel genetic markers can be obtained when the amount of DNA template is as low as 62.5pg; it has species specificity and can only effectively amplify human-derived DNA. increase; high accuracy; suitable for DNA typing of multiple types of inspection materials; the cumulative individual recognition efficiency (Combined power of discrimination, CPD) of 27 autosomal InDel genetic markers of the present invention in the Han population reaches more than 0.999999, Combined power of exclusion induos and trios (CPE) d and CPE t ) were 0.955118 an...

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Abstract

The invention relates to a composite amplification system for jointly detecting InDel genetic markers of human full locus group, and a kit thereof. Particularly, the system jointly detects 45 Insertion/ Deletion (InDel) polymorphic genetic markers on human autosomes and sex chromosomes. The composite amplification system comprises 45 pairs of primers marked by four fluorescent dyes (FAM, HEX, TAMRA and ROX) respectively, and can be used for composite amplification and detection of 45 InDel genetic markers in the same system. The kit can be used for detecting human biological samples and has the characteristics of high sensitivity, good repeatability, strong applicability and the like. 27 autosome InDel markers and 16 X chromosome InDel markers have high cumulative individual recognition rate and proper cumulative non-parent exclusion probability in unrelated individuals of Han nationality, and two Y chromosome InDel markers can meet the requirement of sex judgment in detection. Therefore, the detection kit can provide an effective supplementary detection means for forensic medicine practice.

Description

technical field [0001] The invention belongs to the technical field of biotechnology and genetics, and in particular relates to a compound amplification system for joint detection of InDel genetic markers of the human whole locus group and its kit and application, which are used to detect human autosomes and sex chromosomes with good Insertion / Deletion (Insertion / Deletion, InDel) molecular genetic markers of genetic polymorphisms, specifically related to 45 InDel genetic markers suitable for forensic research applications. Background technique [0002] At present, the main genetic marker used in forensic genetic testing is polymorphic short tandem repeat (STR), which has defects such as high mutation rate, long PCR amplification fragment, and limited number of available markers. Single nucleotide polymorphism (single nucleotide polymorphism, SNP), as a third-generation genetic marker, is a DNA sequence polymorphism caused by a single nucleotide variation at the genome level....

Claims

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Application Information

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IPC IPC(8): C12Q1/6888
CPCC12Q1/6888C12Q2600/156
Inventor 张素华李成涛陶瑞旸张静怡盛翔
Owner ACADEMY OF FORENSIC SCIENCE
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