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A low-frequency and high-penetrance genetic marker related to the auxiliary diagnosis of primary lung cancer and its application

A genetic marker, a technology for assisting diagnosis, applied in the determination/examination of microorganisms, biochemical equipment and methods, DNA/RNA fragments, etc.

Active Publication Date: 2017-06-06
NANJING YIKE POPULATION HEALTH RES INST CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] However, there is no report on the application of low-frequency and high-penetrance genetic markers in the diagnosis of lung cancer. If the low-frequency and high-penetrance genetic variants of lung cancer susceptibility can be screened out as biomarkers, and corresponding diagnostic kits can be developed, it will be beneficial to the diagnosis of lung cancer. The current situation of lung cancer diagnosis in my country will be a powerful impetus, and it will also open up new ways for drug screening, drug efficacy evaluation and targeted therapy

Method used

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  • A low-frequency and high-penetrance genetic marker related to the auxiliary diagnosis of primary lung cancer and its application
  • A low-frequency and high-penetrance genetic marker related to the auxiliary diagnosis of primary lung cancer and its application
  • A low-frequency and high-penetrance genetic marker related to the auxiliary diagnosis of primary lung cancer and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0098] The collection of embodiment 1 sample and the arrangement of sample data

[0099] The inventor collected a large number of blood samples from patients with primary lung cancer from the First Affiliated Hospital of Nanjing Medical University and Jiangsu Cancer Hospital from April 2005 to December 2011. After sorting out the sample data, the inventor selected 4336 experimental samples of whole exome microarray scanning and single genetic marker Sequenom MassARRAY genotyping of samples meeting the following criteria:

[0100] 1. Patients with primary lung cancer diagnosed by histopathology;

[0101] 2. Healthy control;

[0102] The demographic data and clinical data of these samples were collected systematically.

Embodiment 2

[0103] Example 2 Whole Exome Scanning of Genetic Markers in Peripheral Blood DNA

[0104] A total of 1386 eligible primary lung cancer patients and 2950 healthy controls were included above. These two groups of people were passed through Illumina Human Exome BeadChip (Illumina Inc., San Diego, CA) chip detection obtained relevant results. The specific steps are:

[0105] 1. Add hemolysis reagent (i.e. lysate, 40 parts) to the peripheral blood stored in the 2ml cryopreservation tube. Dilute the TrisHcl solution to 2000ml, the same below), turn it upside down and mix it completely.

[0106] 2. Removal of red blood cells: Fill the 5ml centrifuge tube to 4ml with hemolysis reagent, mix by inverting, centrifuge at 4000rpm for 10 minutes, and discard the supernatant. Add 4ml of hemolysis reagent to the precipitate, invert and wash again, centrifuge at 4000rpm for 10 minutes, and discard the supernatant.

[0107] 3. Extract DNA: add 1ml extract solution to the precipitate (eac...

Embodiment 3

[0117] Example 3 Sequenom MassARRAY Genotyping of Single Genetic Markers

[0118] The genetic markers associated with the onset of primary lung cancer found by the above-mentioned whole exome scan were detected on another 1500 primary lung cancer cases and 3000 healthy controls through the Sequenom MassARRAY genotyping platform. The specific steps are as follows:

[0119] 1. Add the hemolysis reagent to the peripheral blood stored in the 2ml cryopreservation tube, mix it upside down and transfer it completely.

[0120] 2. Removal of red blood cells: Fill the 5ml centrifuge tube to 4ml with hemolysis reagent, mix by inverting, centrifuge at 4000rpm for 10 minutes, and discard the supernatant. Add 4ml of hemolysis reagent to the precipitate, invert and wash again, centrifuge at 4000rpm for 10 minutes, and discard the supernatant.

[0121] 3. Extract DNA: Add 1ml of extract solution and 8μl of proteinase K to the precipitate, shake and mix well on the shaker, and bathe in 37°C w...

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Abstract

The invention belongs to the field of genetic engineering and oncology, and discloses a low-frequency high-penetrance genetic marker associated with the auxiliary diagnosis of primary lung carcinoma and applications of the marker. The marker is a combination of 35 genetic loci such as rs1131897, rs1133833, rs115019901, rs117855259, rs118154190, rs141059768, rs141350740, rs142881943, rs145529151, rs145638335, rs145940981, rs150279473, rs150695913, rs151223396, rs185832840 and rs188159138. The marker can be used for preparing kits for the auxiliary diagnosis of primary lung carcinoma.

Description

[0001] field of invention [0002] The invention belongs to the fields of genetic engineering and tumor medicine, and relates to a low-frequency and high-penetrance genetic marker related to auxiliary diagnosis of primary lung cancer and its application. Background technique [0003] Lung cancer is currently the most malignant tumor affecting the number of people worldwide. According to estimates by the International Agency for Research on Cancer (IARC), there were 1.825 million new cases of lung cancer and 1.59 million deaths in 2012, ranking first in the incidence and mortality of cancer worldwide. Due to the continuous growth of tobacco consumption, environmental pollution and changes in lifestyle, the incidence and death of lung cancer in my country have been on the rise in the past two decades. Only from 2000 to 2005, the number of lung cancer cases and deaths due to lung cancer in China increased respectively. It is estimated that in 2012, the incidence rate of lung canc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/68
Inventor 沈洪兵胡志斌靳光付马红霞戴俊程孙洁
Owner NANJING YIKE POPULATION HEALTH RES INST CO LTD
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