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Composition for lung cancer screening and application of composition

A composition and technology for lung cancer, applied in the determination/testing of microorganisms, biochemical equipment and methods, etc., can solve the problems of inability to diagnose early, low sensitivity, high rate of missed diagnosis, etc., to achieve increased sensitivity, ensure correctness and reliability Effect

Inactive Publication Date: 2016-09-07
BIOCHAIN BEIJING SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, compared with prostate cancer and breast cancer, lung cancer still lacks ideal early auxiliary diagnosis methods
[0004] Currently, the lung cancer screening and diagnosis methods used clinically mainly rely on low-dose spiral CT and chest X-ray imaging data examination. In addition, there are many other auxiliary methods to help support the diagnosis of lung cancer, such as fiberoptic bronchoscopy and sputum cytology. However, after long-term research, it has been found that the above-mentioned methods have their own advantages and disadvantages, and they all have the limitations of high cost, large investment, low sensitivity, high missed diagnosis rate, high false positive rate, and inability to make early diagnosis. sex, and some cause pain and inconvenience to patients

Method used

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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0211] Example 1 Primer and Probe Screening

[0212] For the three genes of SHOX2, PTGER4 and FOXL2, many combinations of probes and primers can be designed, and the performance of each combination of probes and primers may be different. Therefore, in the following examples, the probes and primers were screened.

[0213] In this example, the primers and probes of SHOX2, PTGER4 and FOXL2 were first screened with artificial methylated templates and unmethylated templates. This embodiment includes the following steps:

[0214] First, design various primers and probes for SHOX2, PTGER4 and FOXL2, as long as they can be equal to, complementary to or hybridize under moderate stringency or stringent conditions : at least 18 nucleotides of 27 and its complement.

[0215] Then, using artificial methylated oligonucleotide sequences and artificial unmethylated oligonucleotide sequences as templates, PCR amplification was performed using different probe and primer combinations. Whe...

Embodiment 2

[0254] Example 2 Methylated DNA multiplex detection technology for SHOX2, PTGER4 and FOXL2

[0255] Tissue samples from 4 cases of lung cancer (L1-L4), 2 cases of colon cancer (CRC1, CRC2), 1 case of gastric cancer (SC) and 1 case of normal person (WBC) were used as the detection objects. Genomic DNA was extracted from the tissues of the above 4 cases of lung cancer, 2 cases of intestinal cancer, 2 cases of lung cancer, 1 case of blood cancer and 1 case of normal person. All cancer samples were from Boercheng Company. Human normal samples were from BioReclamationIVT Company. The DNA extraction can be performed by any standard means in the prior art. Specifically, in this example, DNA from all human samples is extracted by using EPi proColon Plasma QuickKit from Epigenomics.

[0256] The genomic DNA sample is then pretreated such that the unmethylated cytosine base at the 5' position is converted to uracil, thymine, or another base that hybridizes differently from cytosine...

Embodiment 3

[0262] Example 3 Sensitivity and specificity of methylated DNA multiplex detection kits for SHOX2, PTGER4 and FOXL2 in lung cancer and normal human plasma

[0263] The plasma samples of 20 cases of normal people and 20 cases of lung cancer patients were used as the detection objects. Cell-free DNA was extracted from each sample. The extraction of DNA can be performed by any standard means in the prior art. Specifically, in this embodiment, all sample DNA is extracted by using EPi proColon Plasma Quick Kit from Epigenomics.

[0264] The DNA sample is then pretreated such that the unmethylated cytosine base at the 5' position is converted to uracil, thymine, or another base that hybridizes differently than cytosine. In this example, this pretreatment was achieved by bisulfite reagent treatment. Modification of bisulfite DNA was performed using the EPi proColon Plasma Quick Kit.

[0265] Then, the above pretreated DNA samples of 20 cases of normal people and 20 cases of lun...

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Abstract

The invention provides a composition for lung cancer screening. The composition consists of nucleotide sequences for detecting a methylation degree in at least one target region of each of the following genes: an SHOX2 gene, a PTGER4 gene and an FOXL2 gene, wherein the target regions are continuous segments which are selected from SEQ ID NO: 25, SEQ ID NO: 26 and SEQ ID NO: 27 and are at least 18 bases in length; and the nucleotide sequences are equivalent, complementary or hybridized in the target regions. With the application of the composition disclosed by the invention, the existence of cell hyperplastic abnormity can be shown by integrating methylation detection results of the SHOX2, the PTGER4 and the FOXL2.

Description

technical field [0001] The invention relates to the field of gene detection, in particular to the screening of gene markers, the combined application of the gene markers and the application of the combination of the gene markers in in vitro auxiliary diagnosis of diseases. Background technique [0002] Cancer is a disease caused by abnormal cell proliferation, and it is also a major public health problem. According to the "2012 China Tumor Registration Annual Report" released by the National Cancer Center, the number of malignant tumors in the national tumor registration area is Lung cancer, followed by colorectal cancer, liver cancer and esophageal cancer; the first cause of death is lung cancer, followed by liver cancer, esophageal cancer and colorectal cancer. The morbidity and mortality of lung cancer in my country have leapt to the first place among all kinds of tumors, and it has become one of the diseases that seriously threaten people's health. [0003] The onset of...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6886C12Q2600/154C12Q2600/16
Inventor 周光朋马竣韩晓亮王建铭
Owner BIOCHAIN BEIJING SCI & TECH
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