Method for analyzing mixed sample DNA (Deoxyribose Nucleic Acid) on basis of INDEL-SNP (INDEL and Single Nucleotide Polymorphism) linkage relationship

A technology of mixed samples and relationship analysis, applied in the field of forensic detection, can solve the problems of detection loss, low polymorphism, and no forensic mixed spots, etc., and achieve the effect of protecting safety, advancing time limit, and avoiding amniotic fluid or chorionic villus puncture

Active Publication Date: 2016-11-09
SHANXI MEDICAL UNIV
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Problems solved by technology

But this technology has the following disadvantages (1) instruments and reagents etc. make the method expensive and require professional analysis, which is difficult to popularize; (2) compared with STR analysis, this method can obtain all information on a DNA chain, and the polymorphism is greatly reduced. Improvement, the sensitivity is improved compared with the STR method (can detect 1% DNA), but there is a certain error rate (generally within 2%), forensic analysis requires accuracy to be the first, any base sequence Any misinterpretation will cause irreparable losses to the prosecution
Therefore, the method needs to be carefully evaluated for forensic analysis.
[0007] 3: SNP analysis: There are many kinds of SNP analysis techniques. Since the polymorphism of a single SNP is much lower than that of STR analysis, direct sequencing or electrophoresis is rarely used to detect mixed spots; INDEL can be regarded as a special SNP. At present, there are corresponding kits in foreign countries. In addition to forensic polymorphism analysis, it is also used to analyze the composition ratio of peripheral blood DNA after bone marrow transplantation, but there is no forensic mixed spot analysis.
Because it uses the information of a single SNP, the polymorphism is low, and there is a certain degree of false positives, the experimental conditions are very strict, and it is rarely used in forensic mixed spot analysis and DNA analysis after bone marrow transplantation

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  • Method for analyzing mixed sample DNA (Deoxyribose Nucleic Acid) on basis of INDEL-SNP (INDEL and Single Nucleotide Polymorphism) linkage relationship
  • Method for analyzing mixed sample DNA (Deoxyribose Nucleic Acid) on basis of INDEL-SNP (INDEL and Single Nucleotide Polymorphism) linkage relationship
  • Method for analyzing mixed sample DNA (Deoxyribose Nucleic Acid) on basis of INDEL-SNP (INDEL and Single Nucleotide Polymorphism) linkage relationship

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Embodiment 1

[0046] A kind of method based on INDEL-SNP linkage relationship analysis mixed sample DNA of the present invention adopts the method of INDEL+SNP to solve the analysis problem of mixed spot, and its process is as follows:

[0047] First, select an INDEL site with a good frequency distribution in the Chinese population (the base difference of INDEL is 2bp-10bp), and there is another SNP site with good polymorphism within 300bp downstream or upstream of the INDEL site (selection criteria HET>3.0). Firstly, a PCR product containing two sites of INDEL+SNP is amplified, wherein the upstream primer is specific for the INDEL site, and the downstream primer is designed to include the SNP, and a total of 8 genotypes will appear in the population such as figure 1 as shown, figure 1 are the 8 genotypes that will appear in the population. The long and short boxes represent IN-type and DEL-type INDEL polymorphisms, respectively, and ovals and triangles represent two polymorphisms with SN...

Embodiment 2

[0081] If a spot evidence is extracted from the on-site bed sheet, 11 ID-SNP types are performed on the spot. It was found that the △Cq of rs397832665 and rs397832994 loci were > 8, and the typing could be determined as LL type and SS type respectively. The △Cq of the remaining 9 sites were all <0.5, and the typing could be judged as LS type. The victim was typed as LL and SS at rs397832665 and rs397832994. There were four suspects A, B, C, and D at the scene. At the rs397832665 locus, their types were LL, LS, SS, and LL. Considering the marks and the victim's type, the suspect B and C can be ruled out. At the rs397832994 locus, the types of suspects A and D were LS and SS, respectively. Combining the spot marks and the victim's typing, suspect A could be ruled out. Among the remaining 9 loci of LS type, taking rs397844074 as an example, the victim's type is SS type, and the suspect D's type is LL type, so suspect D cannot be ruled out.

[0082] Analytical method described...

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Abstract

The invention relates to a method for analyzing mixed sample DNA (Deoxyribose Nucleic Acid) on the basis of an INDEL-SNP (INDEL and Single Nucleotide Polymorphism) linkage relationship, and belongs to the technical field of forensic detection. The method comprises: (1) carrying out RT-PCR (Reverse Transcription-Polymerase Chain Reaction) amplification on the mixed sample DNA to obtain a PCR product containing two linkage sites of INDEL and SNP; (2) carrying out parting on the SNP contained in the amplification product obtained in the step (1). The method disclosed by the invention has the following advantages that: a designed INDEL-SNP primer has the species specificity, is suitable for forensic detection, and can be used for DNA detection analysis of forensically common mixed stains no matter whether the mixed strains are formed by the cells of a same type or the cells of different types; the method also can be used for monitoring transplant ingredients of peripheral blood after a bone marrow transplantation operation or other organ transplantation operations, and identification time limit is also advanced greatly.

Description

technical field [0001] The invention belongs to the technical field of forensic medicine detection, and in particular relates to a method for analyzing mixed sample DNA based on INDEL-SNP linkage relationship. The analysis of mixed sample DNA (that is, mixed spots) includes confirming mixed spots and performing individual identification on mixed spots. Background technique [0002] Mixed samples of two or more individuals can be seen in the following situations: (1) Forensic mixed samples include mixed spots of blood samples, saliva spots, semen spots, vaginal secretions, etc. from multiple individuals; (2) patients undergoing bone marrow transplantation Or peripheral blood samples or organs after organ transplantation; (3) detection of early mutations in tumor patients or heterogeneity of tumors; (4) fetal samples mixed in peripheral blood of chimeric individuals or pregnant women, etc. With the development of DNA detection technology, the detection sensitivity of DNA mixed...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6888C12Q2600/156C12Q2565/125C12Q2565/102
Inventor 张更谦贠克明刘晋玎
Owner SHANXI MEDICAL UNIV
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