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A methylation marker for early detection of lung cancer and its detection method

A technology for methylation markers and early detection, applied in biochemical equipment and methods, microbiological determination/inspection, DNA/RNA fragments, etc. Effects of improving quality of life, sensitivity and specificity, and prolonging survival

Active Publication Date: 2019-02-01
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these methods have disadvantages such as cumbersome operation, high false positive rate, and high cost, which limit their wide application in clinical laboratories.

Method used

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  • A methylation marker for early detection of lung cancer and its detection method
  • A methylation marker for early detection of lung cancer and its detection method
  • A methylation marker for early detection of lung cancer and its detection method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] A set of specific primers designed for methylation markers used in the diagnosis of lung cancer:

[0033] According to the whole human genome sequence released by NCBI (National Center for Biotechnology Information), it was designed using PrimerPremier3.0 and Methyl Primer Express v1.0, and synthesized by Shanghai Sangong Co., Ltd. The upstream primer and downstream primer of the PCDHGB6 gene are respectively shown in SEQ ID NO.1 and SEQ ID NO.2; the upstream primer and downstream primer of the HOXA9 gene are respectively SEQ ID NO.3 and SEQ ID NO. 4; the upstream primer and the downstream primer of the MGMT gene are respectively shown in SEQ ID NO.5 and SEQ ID NO.6; the upstream primer and the downstream primer of the MicroRNA-126 gene are respectively SEQ ID NO. 7 and shown in SEQ ID NO.8.

Embodiment 2

[0035] Application of the kit of the present invention in detecting the methylation level of human lung cancer:

[0036] The Master used in the present invention was purchased from Roche Co., Ltd.; the DNA extraction kit was QIAmp DNA MiniKit (QIAGEN); the transformation kit was EZ DNA Methylation kit (Zymo Research, Orange, CA, USA); other reagents were domestic analysis Pure reagents.

[0037] The biological materials used in the present invention are all from Shanghai Zhongshan Hospital.

[0038] method:

[0039] 1. Biological samples:

[0040] Tumor tissues and paired normal tissues of 54 lung cancer patients.

[0041] 2. Extraction and transformation of tissue DNA:

[0042] Take about 25mg of lung cancer tissue or normal tissue, cut it into pieces with surgical scissors after autoclaving and put it into a 1.5ml centrifuge tube, use the DNA extraction kit to extract the DNA of the sample to be tested, and then use the transformation kit to transform the extracted DNA . ...

Embodiment 3

[0055] Operating characteristic curve analysis (ROC curve analysis):

[0056] A ROC curve was constructed to compare the diagnostic ability of the four methylation markers in distinguishing tumor tissue from normal tissue in patients with lung cancer. The areas under the ROC curve of the four markers were as follows: PCDHGB6, 0.796; HOXA9, 0.694; MGMT, 0.594; Mir-126, 0.658. Under the optimal cutoff value, the sensitivity and specificity of the genes are as follows: PCDHGB6, 66.7% and 90.7%; HOXA9, 42.6% and 96.3%; MGMT, 25.9% and 94.4%; Mir-126, 38.9% and 90.7% . The combined detection AUC of these four markers reached 0.891, and the sensitivity and specificity were 85.2% and 81.5% respectively (see Table 3). These results indicated that the combined detection of lung cancer by the four methylation markers has relatively high sensitivity and specificity.

[0057] Table 3 ROC curve analysis of methylation markers

[0058]

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Abstract

The invention relates to a methylation marker for early detection of cancer and a detection method thereof. The methylation marker comprises PCDHGB6, HOXA9, MGMT and microRNA-126 genes. The methylation marker has the advantages of quickness, high throughput, high sensitivity and high specificity when being adopted for combined detection, and the detection method is suitable for early detection of cancer, can be used for late detection of lung cancer and has a good application prospect.

Description

technical field [0001] The invention belongs to the field of lung cancer detection, in particular to a methylation marker for early detection of lung cancer and a detection method thereof. Background technique [0002] Lung cancer, also known as primary bronchial cancer, is one of the most harmful malignant tumors to human health, and its mortality rate ranks first among all kinds of malignant tumors. The World Health Organization predicts that by 2025, there will be more than one million new cases in China every year, which will greatly endanger the lives and health of people in our country. According to research, the overall 5-year survival rate of lung cancer is only 10%-16%, and the 5-year survival rate of patients with stage I can reach 65-75%, while the overall 5-year survival rate of patients with stage II~IV drops from 40% to 10%. 5%. Due to the extremely complex etiology and pathogenesis of lung cancer, and individual differences in clinical manifestations, most p...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6886C12N15/11
CPCC12Q1/6886C12Q2600/154
Inventor 张宏莲刘芳铭毛红菊周麟卢韶华马天乐张其清金庆辉赵建龙
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI