CN-119287026-B - Primer group and kit for identifying hair shaft individuals and application of primer group and kit

Abstract

The invention belongs to the technical field of forensics, and particularly relates to a primer set and a kit for identifying a hair shaft individual and application of the primer set and the kit. The primer group can realize the detection of the cSNP and/or MH locus on Mao Gangao expressed mRNA marks, solves the problem that the hair shaft sample cannot be identified individually, and has higher accuracy. Meanwhile, the instruments and the reagents used in the invention are prepared conventionally in forensic laboratories, and the method is simple and convenient to operate and easy to popularize.

Inventors

• ZHANG GENGQIAN
• LIU JINDING
• YAN JIANGWEI
• YUAN KEMING
• ZHANG YUXIN

Assignees

• 山西医科大学

Dates

Publication Date

20260508

Application Date

20241031

Claims (4)

1. 1. The application of the reagent for detecting the cSNP loci of the human genes in the polymorphism detection of mRNA identified by the hair shaft individual or forensic identification of the hair shaft detection material comprises 404 cSNP loci shown in the following table: ; The reagent comprises a primer group for amplifying the 404 cSNP loci; The primer group comprises 326 pairs of primer pairs, and the nucleotide sequence of the 326 pairs of primer pairs is shown as SEQ ID NO. 1-SEQ ID NO. 652.
2. 2. The use according to claim 1, wherein the reagent further comprises an RNA extraction reagent and/or an RNA reverse transcription reagent.
3. 3. A method for detecting mRNA polymorphisms identified by individuals on the hair shaft, comprising the steps of: Extracting RNA of a hair shaft sample to be detected, and carrying out reverse transcription on the obtained hair shaft sample RNA to obtain hair shaft sample cDNA; Performing multiplex PCR amplification on the hair shaft sample cDNA by using a primer set to obtain a multiplex PCR amplification product of the hair shaft sample, wherein the primer set comprises 326 pairs of primer pairs, and the nucleotide sequences of the 326 pairs of primer pairs are shown as SEQ ID NO. 1-SEQ ID NO. 652; Carrying out library construction based on multiple PCR amplification products of the hair shaft sample to obtain a hair shaft sample sequencing library; performing high-throughput sequencing on the hair shaft sample sequencing library to obtain a high-throughput sequencing result; genotyping based on the high throughput sequencing results to obtain snp and/or microsotype results.
4. 4. The method for detecting an mRNA polymorphism according to claim 3, wherein the threshold value of the number of amplicon reads at the time of genotyping is set at 30.

Description

Primer group and kit for identifying hair shaft individuals and application of primer group and kit Technical Field The invention belongs to the technical field of forensics, and particularly relates to a primer set and a kit for identifying a hair shaft individual and application of the primer set and the kit. Background Hair samples are one of the common samples in forensic identification work, and forensics often collect hair samples at the site of the case, but since hair is mostly naturally shed, hair follicles are multiple or there are no hair follicles, which easily results in failure of detection or insufficient nuclear DNA. Since the hair shaft also contains a certain amount of DNA, although the DNA content in the hair shaft is very small and degraded, there are some methods of individual identification with the hair shaft in the prior art, such as an autosomal Short Tandem Repeat (STR) assay, a mitochondrial DNA assay, and a single amino acid polymorphism assay. However, due to the present situation of trace degradation of the hair shaft DNA, the autosomal STR detection method often has the phenomenon that the alleles of the fragment loci of the long fragment loci are lost or inserted, the difference of single nucleotide polymorphism can be obtained by detecting 2 hypervariable regions of mitochondrial DNA, but the mitochondrial DNA belongs to maternal inheritance and can only be used for eliminating individuals of different maternal lines, and the single amino acid polymorphism method can reversely push the non-synonymous single nucleotide polymorphism of the genomic DNA sequence through the change of the single amino acid polymorphism, but the synonymous mutation on the DNA cannot be reflected on the single amino acid, the information quantity is lower, and the quality deviation during mass spectrometry is caused by the chemical modification or biological modification of the amino acid, so that the error of deducing the non-synonymous single nucleotide polymorphism can be caused. As can be seen, the existing methods for identifying individuals by using hair shafts have some defects which cannot be overcome, and the field is also lack of new genetic markers and detection methods for realizing accurate individual identification of hair shaft samples. Disclosure of Invention The invention aims to provide a primer group for identifying a hair shaft individual, a kit and application thereof, wherein the primer group and the kit containing the primer group can realize detection of cSNP and/or MH sites on Mao Gangao expressed mRNA marks, solve the problem that the hair shaft sample cannot identify the individual, and have higher accuracy. The invention provides application of a reagent for detecting a human gene cSNP locus in one or more of mRNA polymorphism detection of hair shaft individual recognition, forensic identification of hair shaft detection materials and forensic detection of hair shaft detection materials, wherein the human gene cSNP locus comprises 404 cSNPs shown in the following table: The invention also provides a primer set for identifying the hair shaft individuals, which comprises 326 pairs of primer pairs, wherein the nucleotide sequences of the 326 pairs of primer pairs are shown as SEQ ID NO. 1-SEQ ID NO. 652. The invention also provides a kit for identifying the hair shaft individuals, which comprises the primer set according to the technical scheme. The invention also provides application of the primer set or the kit in one or more of mRNA polymorphism detection for hair shaft individual identification, forensic identification of hair shaft detection materials and forensic detection of hair shaft detection materials. The invention also provides an mRNA polymorphism detection method for detecting the recognition of the hair shaft individual, which comprises the following steps: Extracting RNA of a hair shaft sample to be detected, and carrying out reverse transcription on the obtained hair shaft sample RNA to obtain hair shaft sample cDNA; Carrying out multiplex PCR amplification on the hair shaft sample cDNA by using the primer set in the technical scheme to obtain a multiplex PCR amplification product of the hair shaft sample; Carrying out library construction based on multiple PCR amplification products of the hair shaft sample to obtain a hair shaft sample sequencing library; performing high-throughput sequencing on the hair shaft sample sequencing library to obtain a high-throughput sequencing result; genotyping based on the high throughput sequencing results to obtain snp and/or microsotype results. The beneficial effects are that: The invention provides application of a reagent for detecting a human gene cSNP locus in one or more of mRNA polymorphism detection of hair shaft individual recognition, forensic identification of hair shaft detection materials and forensic detection of hair shaft detection materials, wherein the human gene cSNP locus comprises 404 cSNPs. On the