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Kit and device for detecting ATRX and KDM5A mutation based on digital PCR technology and application

A technology for ATRXE17M1571V and mutation detection, applied in the field of biology, can solve problems such as long detection cycle, high cost, and complicated sequencing process, and achieve high sensitivity and accuracy

Active Publication Date: 2020-12-22
BEIJING NOVOGENE TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The first-generation sequencing has a long detection cycle and a sensitivity of about 10%; while the next-generation sequencing process is complex, expensive, and has a long detection cycle and a sensitivity of about 1%.

Method used

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  • Kit and device for detecting ATRX and KDM5A mutation based on digital PCR technology and application
  • Kit and device for detecting ATRX and KDM5A mutation based on digital PCR technology and application
  • Kit and device for detecting ATRX and KDM5A mutation based on digital PCR technology and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment approach

[0042] According to a typical embodiment of the present invention, a kit for detecting ATRX and KDM5A mutations based on digital PCR technology is provided. The kit includes: ATRX E10 del, ATRX E11 del, ATRX E9 Ins, ATRXE14R1426L, ATRX E17 M1571V, ATRX E28 E2074G, KDM5A E27 del, KDM5A E23 P1237S, KDM5A E23 R1217W, KDM5A E19 del, KDM2K E10A Primers and probes for del loci.

[0043] Apply the technical scheme of the present invention, for ATRX E10 del, ATRX E11 del, ATRX E9 Ins, ATRXE14 R1426L, ATRX E17 M1571V, ATRX E28 E2074G, KDM5A E27 del, KDM5A E23 P1237S, KDM5A E23 R1217W, KDM5A E19 del 1, KDM5A E The detection of the set of 12 specific sites of KDM5A E25 del can better reflect the mutation of ATRX and KDM5A. In addition, digital PCR can be used for accurate absolute quantitative detection without relying on control samples and standard curves , to directly detect the copy number of the target sequence, and this detection method has a high sensitivity and precision of abou...

Embodiment 1

[0058] In the examples of the present invention, the experimental methods used are conventional methods unless otherwise specified. The materials and reagents used can be obtained from commercial sources unless otherwise specified.

[0059] The operation steps are as follows:

[0060] 1) Plasma separation and extraction of cfDNA

[0061] Nucleic acid extraction was performed using a commercial cfDNA extraction kit, and DNA concentration was measured with Qubit.

[0062] 2) Prepare digital PCR reaction system

[0063] Using the cfDNA extracted in step 1) as a template, add 1 tube of primer-probe master mix and digital PCR master mix in the above-mentioned kit to prepare a reaction system.

[0064] 3) Chip loading

[0065] Spread the reaction system on the chip to form about 20,000 independent micro-reaction systems.

[0066] 4) PCR amplification

[0067] Put the chip into the PCR amplification instrument symmetrically, set the reaction program, and start the amplification...

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Abstract

The invention discloses a kit and device for detecting ATRX and KDM5A mutation based on a digital PCR technology and application. The kit comprises primers and probes for ATRX E10 del, ATRX E11 del, ATRX E9 Ins, ATRX E14 R1426L, ATRX E17 M1571V, ATRX E28 E2074G, KDM5A E27 del, KDM5A E23 P1237S, KDM5A E23 R1217W, KDM5A E19 del, KDM5A E10 del and KDM5A E25 del loci. By the technical scheme of the invention, detection is carried out for a set of the 12 specific loci, and ATRX and KDM5A mutation conditions can be well reflected.

Description

technical field [0001] The invention relates to the technical field of biology, in particular to a kit, device and application for ATRX and KDM5A mutation detection based on digital PCR technology. Background technique [0002] As the number one enemy of human health, cancer is one of the most important non-communicable diseases in the world, and it is also the chronic disease with the highest fatality rate. According to my country's "2018 National Cancer Report", China added 3.804 million new cancer cases in 2017, which is equivalent to an average of more than 10,000 people being diagnosed with cancer every day, and 7 people are diagnosed with cancer every minute. [0003] Clinically, tissue biopsy is the gold standard for tumor detection. However, due to the dependence on imaging, invasive procedures, difficult operability, and low repeatability in obtaining samples, liquid biopsy technology emerged as the times require. Liquid biopsy has the advantages of non-invasive, e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6886C12Q1/6851
CPCC12Q1/6886C12Q1/6851C12Q2600/156C12Q2531/113C12Q2563/107C12Q2527/101
Inventor 王莹莹成岗杨晶李东东
Owner BEIJING NOVOGENE TECH CO LTD
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