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DNA methylation qPCR kit for lung cancer detection and using method thereof

A technology of methylation and kit, which is applied to the DNA methylation qPCR kit for lung cancer detection and its application field, and can solve the problems of low specificity and sensitivity of the detection method

Inactive Publication Date: 2019-10-29
深圳市新合生物医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0016] The object of the present invention is to provide a blood polygene joint detection primer probe combination kit for early diagnosis, detection or screening of lung cancer, so as to solve the defects of low specificity and sensitivity of detection methods in the prior art

Method used

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  • DNA methylation qPCR kit for lung cancer detection and using method thereof
  • DNA methylation qPCR kit for lung cancer detection and using method thereof
  • DNA methylation qPCR kit for lung cancer detection and using method thereof

Examples

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Effect test

Embodiment 1

[0058] Example 1: Using the kit of the present invention to perform multiple qPCR detection of lung cancer methylation on healthy human cfDNA samples

[0059] (1) Experimental materials

[0060] 1. Healthy human plasma (purchased from Bloodworks NW);

[0061] 2. QIAamp Circulatory System Nucleic Acid Extraction Kit (purchased from Qiagen, Cat. No. 55114);

[0062] 3. EZ DNA Methylation-Lightning Kit (purchased from Zymo Research, Cat. No. D5031);

[0063] 4. The kit of the present invention (which includes a set of primers and probe combinations for detecting the targets SHOX2 and SOX17, ACTB amplification primers and probes);

[0064] 5. AmpliTaq Gold TM DNA polymerase and buffer reagents (purchased from Thermo Fisher, Cat. No. 4311806).

[0065] (2) Experimental method

[0066] 1. DNA extraction from plasma

[0067] cfDNA was extracted from 5 healthy human plasma samples using the QIAamp Circulatory System Nucleic Acid Extraction Kit, and the steps were performed acco...

Embodiment 2

[0080] Example 2: Using the kit of the present invention to perform multiple qPCR detection of lung cancer methylation on ctDNA samples from lung cancer patients

[0081] (1) Experimental materials

[0082] 1. Plasma from lung cancer patients (purchased from Bloodworks NW);

[0083] 2. QIAamp Circulatory System Nucleic Acid Extraction Kit (purchased from Qiagen, Cat. No. 55114);

[0084] 3. EZ DNA Methylation-Lightning Kit (purchased from Zymo Research, Cat. No. D5031);

[0085] 4. The kit of the present invention (which includes a set of primers and probe combinations for detecting the targets SHOX2 and SOX17, ACTB amplification primers and probes);

[0086] 5. AmpliTaq Gold TM DNA polymerase and buffer reagents (purchased from Thermo Fisher, Cat. No. 4311806).

[0087] (2) Experimental method

[0088] 1. DNA extraction from plasma

[0089] ctDNA was extracted from 5 plasma samples of lung cancer patients using the QIAamp Circulatory System Nucleic Acid Extraction Kit,...

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Abstract

The invention belongs to the technical field of biotechnologies and DNA detection, and particularly relates to a DNA methylation qPCR kit for lung cancer detection and a using method thereof, in particular to a kit using plasma free DNA to conduct methylation of qCPR to determine the methylation states of one or more gene targets (such as SHOX2 and SOX17) for lung cancer detection or screening anda using method of the kit. The kit can significantly increase the positive detection rate of early lung cancer and improve specificity of the early lung cancer through an experimental test, detectionsensitivity of biomarkers can be improved to picogram / nanogram level DNA molecules, and improvement of the detection sensitivity is realized by optimizing specific nucleotide sequence primers and DNAprobes and improving a hydrosulphite treatment method of the plasma free DNA.

Description

technical field [0001] The invention belongs to the technical field of biotechnology and DNA detection, and in particular relates to a DNA methylation qPCR kit for lung cancer detection and its application method, in particular, to a methylation qPCR using plasma free DNA to determine The methylation status of one or more target gene targets (such as SHOX2 and SOX17) is used for lung cancer detection or screening kits and methods of use thereof. In addition, the present invention also relates to the application of the above detection kits in biomedicine application. Background technique [0002] Lung cancer is one of the most common malignant tumors in humans, and its mortality rate ranks first in cancer deaths. According to the American Cancer Society, there were approximately 234,030 new lung cancer cases and 154,050 lung cancer deaths in the United States alone in 2018, and the estimated number of lung cancer deaths is roughly equivalent to colon cancer, breast cancer, p...

Claims

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Application Information

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IPC IPC(8): C12Q1/6886C12Q1/6858C12N15/11
CPCC12Q1/6886C12Q1/6858C12Q2600/154C12Q2600/166C12Q2531/113C12Q2563/107C12Q2523/125C12Q2545/101
Inventor 王志强万季潘晓新夏迪余涛金泰庆罗海燕蔡雪儿宋麒
Owner 深圳市新合生物医疗科技有限公司
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