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Multiplex amplification kit for simultaneously detecting autosome and Y chromosome STR loci

A compound amplification and Y chromosome technology, applied in the field of compound amplification kits for simultaneous detection of autosome and Y chromosome STR loci, can solve the problem of shortening detection time, detection failure, detection time delay and the best chance to capture criminal suspects and other problems to achieve the effect of shortening the detection time and saving the amount

Pending Publication Date: 2019-08-23
苏州市公安局刑事科学技术研究所 +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At the same time, the STR typing technology is affected by factors such as the state of the test material, the number of amplification sites, and the amplification system. Only the PCR process takes about 3 hours, which often delays the arrest of criminals due to the increased detection time. The best chance for the suspect, but also due to the small amount of physical evidence left by the suspect at the scene, the detection will fail, and eventually the clues will be lost
[0004] All in all, with the increase of criminal cases, the existing detection kits can no longer meet the requirements of public security for the rapid detection and on-site detection of forensic DNA in actual combat. How to realize the simultaneous detection of autosomal STR loci and Y chromosome STR loci , and the ability to shorten the detection time while improving the efficiency of excluding and identifying criminal suspects has become a problem to be solved

Method used

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  • Multiplex amplification kit for simultaneously detecting autosome and Y chromosome STR loci
  • Multiplex amplification kit for simultaneously detecting autosome and Y chromosome STR loci
  • Multiplex amplification kit for simultaneously detecting autosome and Y chromosome STR loci

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Screening of autosomes and Y chromosome STR loci

[0041] The screening of the locus of the present invention is based on the locus disclosed in Chinese Patent Application No. 201010282414.X, and the D6S1043 locus is changed to the D19S433 locus, and the genotype detection of more than 6,000 individuals is performed on the remaining euchromatic loci According to the distribution frequency of alleles of each locus, data such as individual recognition ability (DP), heterozygosity (H), and non-parent exclusion rate (PE) are calculated, wherein, the above data show that in the 16 STR loci, except Except for the TH01 and TPOX loci, the DP values ​​of the other loci are all close to 0.9, the H values ​​are all greater than 0.7, and the PE values ​​are mostly above 0.5, which indicates that the other loci except the TH01 and TPOX loci have good forensic Value. Based on this, the present invention maintains the original autosomal STR loci, and through the identificat...

Embodiment 2

[0046] Embodiment 2 Specific primer design and primer validation

[0047] With the increase of the number of primers in the multiplex amplification system, the mutual interference between primers at different loci becomes more and more serious, and the kinetics of the reaction system become more and more complex, so it is necessary to design a large number of primer sequences for complex testing And explore the specific concentration ratio between the primers in the compound amplification system, and finally ensure that more human autosomes and Y chromosome STR loci are compounded without reducing the specificity and sensitivity of the kit.

[0048] Specifically, the following steps are included:

[0049] (1) Specific primer design

[0050]Before designing primers, it is necessary to analyze the properties of the target sequence to be tested, and select a highly conserved region with uniform base distribution for primer design; in addition, in addition to polymorphisms contai...

Embodiment 3

[0063] The determination of embodiment 3PCR amplification system

[0064] In addition to the specific amplification primers for the above loci, the reaction system of this kit also includes the reaction mixture, hot start Taq enzyme and sdH 2 O, the reaction mixture contains Tris-HCl, MgCl 2 , KCl, dNTP, BSA, the concentration of each component can be adjusted as needed.

[0065] Specifically, in the PCR amplification system, for Mg 2+ The two factors of concentration and dNTPs concentration were optimized by designing an orthogonal experiment, and prepared into a PCR reaction mixture, which was added to the PCR system. The components of the final PCR system are shown in Table 4.

[0066] Table 4 PCR reaction amplification system

[0067]

[0068]

[0069] Wherein, the reaction final concentration of each component in the reaction mixture is 50mM Tris-HCl, 50mM KCl, 2.0mM MgCl 2 , 0.2mM dNTPs, 0.8mg / mL BSA; the primers and primer concentrations corresponding to the ...

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Abstract

The invention discloses a multiplex amplification kit for simultaneously detecting autosome and Y chromosome STR loci. The kit contains 36 groups of specific amplification primer pairs corresponding to 38 loci, and comprises sequences SEQ ID NO. 1-74. A-STR data and Y-STR data can be simultaneously analyzed and used for rapid screening of cases, so that the detection time is shortened, and meanwhile the efficiency of excluding and determining criminal suspects is improved.

Description

technical field [0001] The invention belongs to the field of analytical genetics, relates to a multiple amplification test system for human chromosome STR loci, in particular to a multiple amplification kit for simultaneously detecting autosome and Y chromosome STR loci. Background technique [0002] Short tandem repeat sequence (STR) is a molecular genetic marker commonly used at present. Its fragment is small and easy to amplify. It is suitable for testing trace and degraded biological samples. The amplification conditions of each locus are similar to achieve multiple amplification and automatic detection. , so it has the advantages of sensitivity, accuracy, speed, and large amount of information, and is very suitable for establishing a DNA database. Therefore, DNA-STR typing technology has become one of the main means of forensic evidence testing. [0003] At present, STR typing for forensic testing includes autosomal STR typing and Y chromosome typing detection, and auto...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6888C12Q1/6844
CPCC12Q1/6888C12Q1/6844C12Q2600/16C12Q2525/151
Inventor 周如华石云杰张健王鑫陈维忠陈晶晶崔扬秦东梅柳勇李发院郭育林夏子芳
Owner 苏州市公安局刑事科学技术研究所
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