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High-throughput sequencing-based detection kit for STR loci and detection method

A detection kit and detection method technology, applied in the field of forensic genetics, can solve the problems of inability to provide STR sequence polymorphism information, reduce system efficiency, etc., and achieve the effect of saving DNA sample volume and detection time, and enriching the amount of information

Active Publication Date: 2017-08-04
ACADEMY OF FORENSIC SCIENCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In addition, capillary electrophoresis is used to classify STR detection results based on fragment size. It only detects length polymorphisms and cannot provide sequence polymorphism information within STRs, reducing system efficiency.

Method used

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  • High-throughput sequencing-based detection kit for STR loci and detection method
  • High-throughput sequencing-based detection kit for STR loci and detection method
  • High-throughput sequencing-based detection kit for STR loci and detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] This example is the allele and sequence structure of the D8S1179 locus obtained from 200 unrelated individuals using this kit.

[0048] Unrelated individual blood spots: provided by volunteers.

[0049] Sample DNA extraction: obtain genomic DNA by silica bead method or other extraction methods.

[0050] Sample test results: The alleles and sequence structures of the D8S1179 locus obtained from 200 unrelated individuals using this kit are shown in the table below:

[0051] loci allele core sequence structure D8S1179 7 [TCTA] 7 D8S1179 8 [TCTA] 8 D8S1179 9 [TCTA] 9 D8S1179 10 [TCTA] 10 D8S1179 11 [TCTA]11 D8S1179 12 [TCTA]1[TCTG]1[TCTA]10 D8S1179 12 [TCTA]12 D8S1179 13 [TCTA]1[TCTG]1[TCTA]11 D8S1179 13 [TCTA]13 D8S1179 13 [TCTA]1[TCTG]1[TCTA]11 D8S1179 14 [TCTA]1[TCTG]1[TCTA]12

[0052] loci allele core sequence structure D8S1179 14 [TCTA]2[TCTG...

Embodiment 2

[0055] This example is the application of the above-mentioned kit in the identification of suspected mutations in triplet relatives.

[0056] A case of triplet kinship testing involving the biological mother-child (female)-suspect father.

[0057] STR detection results based on CE technology: 19 autosomal STR loci were detected using the conventional PCR-CE detection kit Goldeye 20A (Beijing Basic Cognitive Biology Co., Ltd.), and it was found that the mother and child were completely identical at the 19 STR loci. According to Mendel's law of inheritance, the suspect father and child had one-step mutations at two STR loci (vWA and FGA). Through calculation, the cumulative parentage index CPI is 378.361. According to the triplet paternity test standard, it is impossible to judge the parent-child relationship between the suspicious father and the child. After background investigation of the parties involved in the case, it was found that the biological mother of the child in th...

Embodiment 3

[0061] This embodiment is the application of the above kit in individual identification of mixed samples.

[0062] This kit is used to detect mixed male-female samples with known mixing ratios of 100:1, 50:1, 10:1 and 1:1. The results show that: when the ratio of the two differs by 50 times, the minor components of the alleles can still be detected; when the mixing ratio increases to 100:1, the information of the minor components of the alleles cannot be effectively extracted (cannot be detected, Or the detection ratio is too low, or it is difficult to distinguish from the Stutter peak). Taking the D2S1776 locus as an example, such as image 3 As shown, the genotype of the female sample is 9-11, and the genotype of the male sample is 10-11, both share the same allele, and the genotyping result of the mixed sample at 50:1 is 9 / 11 / [10], and can be measured according to the read depth (Reads) of high-throughput sequencing. In addition, when two contributors are found to type ...

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Abstract

The invention relates to a high-throughput sequencing-based detection kit for STR loci. The detection kit is used for amplifying 34 STR loci and a sex locus. The kit comprises a primer composition formed by primer pairs of the 34 STR loci and a sex locus, wherein the sequence of the primer composition is as shown in SEQ ID NO:1 to SEQ ID NO:70. The invention further provides a detection method using the kit, and an application of the kit in the field of triplet paternity test, diad paternity test, grandparent and grandchild test, siblings identification and individual recognition. Parallel sequencing is carried out on the STR loci by adopting a high-throughput sequencing technology, and variation information in the sequence and variation information of a flanking sequence can be obtained while allele typing information is read; sequence information of 384 samples on the 34 STR loci can be obtained through single sequencing, so that the DNA sample capacity is reduced and the detection time is shortened; and fragment libraries are concentrated at 150-300bp and more abundant information amount can be obtained for degradable materials.

Description

technical field [0001] The invention belongs to the field of forensic genetics and relates to the detection of STR genetic markers with good forensic application value in human genome, in particular to a detection kit for 34 STR loci based on high-throughput sequencing and its application and detection method. Background technique [0002] STR loci, also known as microsatellite DNA, widely exist in prokaryotic and eukaryotic genomes, accounting for about 3% of the entire genome, and are a class of dozens of nucleosides consisting of 2 to 6 nucleotides as repeating units Acid repeats. The polymorphism of the STR locus is mainly derived from the difference in the number of core sequence repeats among individuals, and this difference follows the Mendelian law of inheritance in the genetic process. Therefore, STR detection technology is widely used in forensic individual identification and kinship identification. [0003] PCR-STR composite fluorescence amplification detection ...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6869C12Q1/6888C12Q2600/156C12Q2525/191C12Q2531/113C12Q2535/122
Inventor 李成涛张素华刘希玲边英男
Owner ACADEMY OF FORENSIC SCIENCE
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