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SNP (Single Nucleotide Polymorphism) marker correlated to assistant diagnosis of primary lung cancer and application thereof

A technology for auxiliary diagnosis and markers, applied in the field of SNP markers

Active Publication Date: 2012-07-04
南京江北新区生物医药公共服务平台有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, there is no report on the application of SNPs in the diagnosis of primary lung cancer. If the SNPs of primary lung cancer susceptibility can be screened out as biomarkers, and corresponding diagnostic kits can be developed, the diagnosis status of primary lung cancer in my country will be improved. It will definitely 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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  • SNP (Single Nucleotide Polymorphism) marker correlated to assistant diagnosis of primary lung cancer and application thereof
  • SNP (Single Nucleotide Polymorphism) marker correlated to assistant diagnosis of primary lung cancer and application thereof
  • SNP (Single Nucleotide Polymorphism) marker correlated to assistant diagnosis of primary lung cancer and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0067] Example 1 The collection of samples and the arrangement of sample data

[0068] The inventor collected a large number of blood samples from patients with primary lung cancer from the Cancer Center of Nanjing Medical University from April 2005 to January 2011. After sorting out the sample data, the inventor selected 9934 samples that met the following criteria Experimental samples for genome-wide microarray scanning and single SNP TaqMan genotyping:

[0069] 1. Patients with primary lung cancer diagnosed by histopathology;

[0070] 2. Healthy controls matched with the age and sex of the cases;

[0071] The demographic data and clinical data of these samples were collected systematically.

Embodiment 2

[0072] Example 2 Whole Genome Scanning of SNPs in Peripheral Blood DNA

[0073] Among the 2,331 primary lung cancer patients and 3,077 healthy controls who met the above criteria, the two groups were matched in age and gender. The two groups of people were detected by Affymetrix6.0 chip to obtain relevant results. The specific steps are:

[0074] 1. Add the hemolysis reagent to the leukocytes stored in the 2ml cryopreservation tube, mix it upside down and transfer it completely.

[0075] 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.

[0076] 3. Extract DNA: Add 1ml extract solution to the precipitate (each 300ml contains 122.5ml 0.2M sodium chloride, 14.4ml 0.5M ethylenediaminetetraacetic acid, 15ml 10% sodium...

Embodiment

[0086] Example 3 TaqMan genotyping of a single SNP

[0087] The SNPs found to be associated with the onset of primary lung cancer in the above genome-wide scan were detected in another 2283 primary lung cancer cases and 2243 healthy controls. The specific steps are as follows:

[0088] 1. Add the hemolysis reagent to the leukocytes stored in the 2ml cryopreservation tube, mix it upside down and transfer it completely.

[0089] 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.

[0090] 3. Extract DNA: Add 1ml of extract solution and 8μl of proteinase K to the precipitate, fully oscillate and mix on a shaker, and bathe overnight at 37°C.

[0091] 4. Remove protein: add 1ml of saturated phenol and mix well (shake gent...

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Abstract

The invention belongs to the fields of gene engineering and tumor medicines, and discloses an SNP (Single Nucleotide Polymorphism) marker correlated to assistant diagnosis of primary lung cancer and application thereof. The marker is a combination of the following 63 SNP: rs107401, rs465498, rs663414, rs622917, rs636586, rs741061, rs855641, rs948783, rs1544694, rs1663689, rs2030378, rs2225588, rs2317683, rs2395166, rs2405271, rs2788713, rs3130975, rs4344933, rs4488809 and the like. The marker can be used for preparing an assistant diagnosis kit for the primary lung cancer.

Description

field of invention [0001] The invention belongs to the fields of genetic engineering and tumor medicine, and relates to a SNP marker related to auxiliary diagnosis of primary lung cancer and its application. Background technique [0002] Lung cancer is currently the most malignant tumor affecting the number of people worldwide. According to the International Agency for Research on Cancer (IARC), there were 1.607 million new cases of lung cancer and 1.375 million deaths in 2008, ranking first in the incidence and mortality of tumors 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 2008, the incidence rate of lung cancer in my country was 45.9 / 100,000 for males and 21.3 / 100,...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 沈洪兵胡志斌马红霞靳光付董静孙重期
Owner 南京江北新区生物医药公共服务平台有限公司
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