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Multiplex amplification kit containing 33 loca of human genome and application of multiplex amplification kit

A composite amplification and locus technology, applied in the field of composite amplification kits, can solve the problems of shortened detection time, failed detection clues, and reduced detection efficiency, and achieves saving manpower, material and financial resources, strong amplification specificity and temperature resistance. receptive effect

Inactive Publication Date: 2016-06-22
GUANGDONG HUAMEI ZHONGYUAN BIOLOGICAL SCI & TECH +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because the existing STR typing technology is affected by factors such as the state of the sample, the amplification system, and the detection throughput, the two-time inspection procedure increases the detection time and often delays the best chance of catching the suspect. The small amount of physical evidence left by the suspect at the scene leads to a decrease in detection efficiency or even failure, which cannot help provide more clues for investigation
Existing detection kits can no longer meet the requirements of public security for rapid detection of forensic DNA and on-site detection capabilities. How to realize the detection of A-STR loci and Y-STR loci together to shorten the detection time and improve the detection efficiency become a problem to be solved

Method used

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  • Multiplex amplification kit containing 33 loca of human genome and application of multiplex amplification kit
  • Multiplex amplification kit containing 33 loca of human genome and application of multiplex amplification kit
  • Multiplex amplification kit containing 33 loca of human genome and application of multiplex amplification kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] The corresponding information of the 33 loci used in the present invention are shown in Table 2 below:

[0041] Table 2. Information of each locus

[0042]

[0043]

[0044] Among the above-mentioned loci, those marked with a / b after the name of the loci are two loci.

[0045] The main components of the detection kit:

[0046] The primer pairs corresponding to the 33 loci are shown in Table 3:

[0047] Table 3. Primer sequences and concentrations for each locus

[0048]

[0049]

[0050]

[0051] The 33 loci were grouped and fluorescently labeled as follows: D3S1358, D13S317, D7S820, D16S539, PentaE, and DYS635 were the first group, and the fluorescent dye marker was 6-FAM; DYS456, TPOX, TH01, D2S1338, CSF1PO, DYS385a / b and DYS458 are the second group, and the fluorescent dye marker is HEX; DYS391, D19S433, vWA, D21S11, D18S51, D6S1043 are the third group, and the fluorescent dye marker is TAMRA; Amelogenin, D8S1179, D5S818, D12S391, FGA, DYS438 is th...

Embodiment 2

[0062] Using the protocol in Example 1, the 14 Y-STR loci in the 33-locus fluorescent-labeled multiplex amplification kit were used for case investigation. A public security bureau provided 1056 samples from different individuals: including 350 blood samples, 330 sperm spots, 260 saliva spots, and 116 tissues. Genomic DNA was extracted with reference to "GA / T383-2014 Forensic Science DNA Laboratory Inspection Specification". Carry out PCR amplification, electrophoresis detection and result analysis according to Example 1. All samples were amplified and analyzed simultaneously with the Yfiler kit, the kit of invention patent 201310364183.0 and the kit of 201410208458.6. The results are shown in Table 6:

[0063] Table 6. Statistics on the number of haplotypes in the 4Y, 10Y, 14Y typing results of 1056 templates

[0064]

[0065]

[0066] With the increase of analysis data, the advantages of the resolution performance of the present invention are obvious. The test resu...

Embodiment 3

[0068] Application of the kit of the present invention in forensic individual identification:

[0069] 1. Collect the blood spots in the case: The sample is the blood spot No. 9948 provided by a public security bureau.

[0070] 2. Sample DNA extraction: The extraction of genomic DNA refers to the "GA / T383-2014 Forensic Science DNA Laboratory Inspection Specification".

[0071] 3. PCR amplification and capillary electrophoresis analysis, the results are as follows: figure 1 , figure 2 , image 3 .

[0072] This kit realizes simultaneous detection and analysis of A-STR loci and Y-STR loci. The inspection procedure that originally required two times is reduced to one completion, which reduces the detection time and improves the detection efficiency, thereby increasing the probability of arresting criminal suspects. Opportunity; another test reduces the absolute amount of samples required, thereby increasing the amount of effective physical evidence left by the suspect at the...

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Abstract

The invention provides a multiplex amplification kit containing 33 loca of a human genome. The multiplex amplification kit contains 18 A-STR loca which are recommended by a DNA database of the Ministry of Public Security to be used and also contains 14 Y-STR loca which are low in mutation rate and high in polymorphism and the Amelogenin locus, simultaneous amplification and detection on the 33 loca through a single tube is achieved, and synchronous amplification and detection on autosomes and Y chromosomes are achieved in a single experiment. The kit can directly amplify blood stains and saliva stains which take filter paper and an FTA filter as carriers without needing the template extraction and purification process and also can be suitable for DNA samples extracted through different extraction methods. The kit can be used for rapid investigation of a case, can improve the detection efficiency and increase the case investigation speed and especially has the significant effect on male sample trace detection in a raping case and father-child paternity test detection. The kit is wide in application range, good in compatibility and completely compatible with an existing legal medical expert DNA detection system.

Description

technical field [0001] The invention relates to a composite amplification kit for simultaneous analysis of human genome autosome and Y chromosome loci, in particular to a composite amplification kit for 33 loci of the human genome and its application. Background technique [0002] Short tandem repeat sequence (STR) is a molecular genetic marker commonly used at present. Its fragments are 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 DNA databases. In the 1990s, the US FBI selected 13 autosomal STR loci (A-STR for short) to establish a DNA data index system - CODIS (Combined DNA Index System): CSF1PO, D3S1358, D5S818, D7S820, D8S1179, D13S317, D16S539, D18S51 , D21S11, FGA, TH01, TPOX, vW...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6888C12Q2600/16C12Q2600/156C12Q2600/124C12Q1/68
Inventor 杜蔚安刘超李发院刘宏张勇果钱水王邦超郑卫国卢青忻星
Owner GUANGDONG HUAMEI ZHONGYUAN BIOLOGICAL SCI & TECH
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