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Composition for noninvasive screening of early lung cancer and application of composition

A composition and lung cancer technology, applied in the direction of microbial measurement/testing, biochemical equipment and methods, etc., can solve the problem of less circulating DNA in tumors, and achieve the effects of easy interpretation, simple operation, and low requirements for instruments

Active Publication Date: 2017-08-11
北京鑫诺美迪基因检测技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the amount of tumor circulating DNA in plasma is small and prone to degradation

Method used

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  • Composition for noninvasive screening of early lung cancer and application of composition
  • Composition for noninvasive screening of early lung cancer and application of composition
  • Composition for noninvasive screening of early lung cancer and application of composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0061] Detection of methylation of SHOX2 and PTGER4 genes in plasma cell-free DNA of patients with stage I lung cancer.

[0062] The components of the kit are as follows:

[0063]

[0064] Plasma samples from 5 healthy individuals and 5 tumor plasma samples with known pathological information as stage I lung cancer were selected.

[0065] 1. Extract free DNA from the above 10 plasma samples.

[0066] 2. After the extraction of free DNA is completed, bisulfite conversion is performed on the extracted DNA, and the unmethylated cytosine (C) is converted into uracil (U), while the methylated cytosine (C) remains unchanged. Purified Bis-DNA was obtained.

[0067] 3. Prepare PCR reaction solution: DNA polymerase, dNTPs, Mg 2+ , 10×DNA polymerase buffer, SHOX2 and PTGER4 gene 1 primer probe, SHOX2 and PTGER4 gene 2 primer probe, SHOX2 and PTGER4 gene 3 primer probe, internal reference gene primer probe, SHOX2 and PTGER4 gene 1 degenerate blocking primer , SHOX2 and PTGER4 gene...

Embodiment 2

[0089] Plasma samples from 93 early lung cancer patients (27 cases at stage 0 and 66 cases at stage I) from Shanghai Lung Hospital and 100 normal human plasma samples from Shandong Qilu Hospital were collected, and SHOX2 and PTGER4 genes were tested for these 193 plasma samples For the detection of methylation, the specific operation steps are as described in Example 1. Among the detection results of 100 normal plasma samples, the difference between the target gene 1 / 2 / 3Ct value and the internal reference gene Ct value in 95 samples was greater than 8, and the specificity reached 95%. For 93 early lung cancer plasma samples, 88 The difference between the target gene 1 / 2 / 3Ct value and the internal reference gene Ct value is less than 8, and its sensitivity reaches 95%, among which 0 stage lung cancer is 89%, and stage I lung cancer is 97%.

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Abstract

The invention discloses a composition for noninvasive screening of an early lung cancer and application of the composition. The composition comprises three pairs of methylation-specific primer probes of SHOX2 (Short Stature Homebox2) genes, three pairs of methylation-specific primers of PTGER4 (Prostaglandin E Receptor 4) genes and one pair of reference gene primer probes and further comprises three MGB probes corresponding to non-methylation target genes in the SHOX2 genes and three MGB probes corresponding to the non-methylation target genes in the PTGER4 genes. A methylation detection kit for the noninvasive screening of the early lung cancer comprises the composition. Patients suffering from the early lung cancer are screened by detecting promoters of the SHOX2 genes and the PTGER4 genes of free DNA in blood of high-risk lung cancer individuals and methylation conditions of three target sites in gene coding and non-coding areas, and early diagnosis of the lung cancer is aided. The methylation-specific PCR / MSP detection method is simple and convenient in operation and low in time consumption, and the results are easily analyzed.

Description

technical field [0001] The invention relates to the technical field of ultra-early screening and detection of lung cancer, in particular to a non-invasive screening technology for stage 0 and stage I of lung cancer based on plasma free DNA. Background technique [0002] According to the latest data from the National Cancer Registry Center, there are about 733,000 new cases of lung cancer in the country, ranking first in the incidence rate of all malignant tumors (19.9%); about 591,000 deaths, accounting for all malignant tumor deaths No. 1 (26.5%). Under the current medical system, the five-year survival rate of patients with advanced lung cancer is only 10-20%, but for the treatment of patients with early-stage lung cancer, about 80-90% of them can be cured by surgical resection. Therefore, early screening of lung cancer is of great significance. Effective early screening and intervention can reduce the incidence of lung cancer by 60%, and the mortality rate by 80%. Howev...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCC12Q1/6858C12Q1/6886C12Q2600/154C12Q2531/113C12Q2545/101C12Q2561/101
Inventor 刘沛
Owner 北京鑫诺美迪基因检测技术有限公司
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