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Reagent kit for detecting lung cancer susceptibility by ERCC2 gene

A kit and susceptibility technology, applied in the field of kits for detecting lung cancer susceptibility, can solve the problems of accumulation of carcinogenic damage, failure to correctly identify DNA damage sites, and failure of base excision repair to proceed correctly.

Inactive Publication Date: 2008-02-27
XINBAXIANG SHANGHAI MOLECULAR MEDICAL TECH SHANGHAI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Loss of function of ERCC2 results in the inability of the essential transcription factor complex to correctly recognize sites of DNA damage, thereby preventing base excision repair from proceeding correctly, leading to the accumulation of oncogenic lesions on DNA

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1. Use of detection kits

[0024] Step 1: Extraction of DNA template

[0025] Genomic DNA of oral epithelial cells was extracted by silica gel adsorption.

[0026] Step 2: Real-time quantitative PCR reaction

[0027] Use a fluorescent quantitative PCR detection kit that can detect susceptibility to lung cancer, which contains the following primer pairs and fluorescent probes:

[0028] Sense primer: 5'-ACTCAGGAGTCACCAGGAA-3' (SEQ ID NO: 2) with a Tm value of 58°C

[0029] Antisense primer: 5'-TCTGTTCTCTGCAGGAGGA-3' (SEQ ID NO: 3) with a Tm value of 58°C

[0030] Fluorescent probe: 5'-AATCTGCTCTTATCCTCTGCAGC-3' (SEQ ID NO: 4) with a Tm value of 66°C

[0031] The fluorescent quantitative PCP reaction system has a total volume of 10 μl, including 2 μl of DNA template with a concentration of 20 ng / μl, 1 μl of 10X fluorescent quantitative PCR reaction buffer, 0.1 μl of 25mM dNTP mixture, and 0.6 μl of 25mM MgCl 2 solution, 0.025 μl (5 units / μl) Taq DNA polymerase...

Embodiment 2

[0035] Example 2. Services that guide people to proactively prevent lung cancer

[0036] At present, Shanghai Zhujian Bioengineering Co., Ltd. has launched this service.

[0037] Step 1: DNA Extraction

[0038] The subjects were consulted before the service, and the subjects signed the informed consent and filled out the questionnaire on living and eating habits. According to the instructions in the sampling box, the oral epithelial cells were sampled with oral swabs, and the DNA of the oral epithelial cells was extracted by silica gel adsorption.

[0039] Step 2: Genotyping Assays

[0040] Using the kit provided by the present invention, the rs13181 SNP site on the ERCC2 gene of the tested subject's DNA is detected by fluorescence quantitative PCR, and the genotype of the SNP site is determined.

[0041] Step 3: Instruct people to be proactive in preventing lung cancer

[0042] Through the analysis of the SNP genotype of the tested person, the genotyping test report and the...

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Abstract

The present invention discloses a kit for detecting pulmonary carcinoma susceptibility. Said kit includes specific primer pair and specific fluorescent probe for detecting SNP site of position 156 in ERCC2 gene SEQ ID NO.1 and conventional component for making fluorescent quantitative PCR detection.

Description

technical field [0001] The present invention relates to a kit for detecting disease susceptibility, especially a kit for detecting susceptibility to lung cancer. Single nucleotide polymorphism (SNP) loci to predict individual susceptibility to lung cancer. Background technique [0002] Primary bronchial lung cancer, referred to as lung cancer, is one of the most common malignant tumors in the world, and it is a disease that seriously threatens people's health and life. Tumor cells originate from the bronchial mucosa or glands, and often have regional lymph node and hematogenous metastasis. In the early stage, respiratory symptoms such as irritating cough and blood in the sputum are often present. The speed of disease progression is related to the biological characteristics of the cells. [0003] For half a century, the morbidity and mortality of lung cancer in all countries in the world have been increasing significantly. The World Health Organization (WHO) reported in 200...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 冯哲民邹祖烨
Owner XINBAXIANG SHANGHAI MOLECULAR MEDICAL TECH SHANGHAI
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