Reagent kit for detecting cancer-related DNA methylation and application of reagent kit for detecting cancer-related DNA methylation

A methylation and gene technology, applied in recombinant DNA technology, DNA/RNA fragment, determination/inspection of microorganisms, etc., can solve problems such as difficult diagnosis, and achieve the effect of good specificity and high diagnostic sensitivity

Active Publication Date: 2015-07-08
INST OF CHEM CHINESE ACAD OF SCI
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with previous statistical methods, this method fully considers the methylation degree and influence of different genes, however, this method requires statistical analysis of a large number of samples, so it is difficult to apply to instant diagnosis of a single sample or a small number of samples

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Reagent kit for detecting cancer-related DNA methylation and application of reagent kit for detecting cancer-related DNA methylation
  • Reagent kit for detecting cancer-related DNA methylation and application of reagent kit for detecting cancer-related DNA methylation
  • Reagent kit for detecting cancer-related DNA methylation and application of reagent kit for detecting cancer-related DNA methylation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1, synthetic PFPN

[0042]Add 1,6-dibromohexane (23 mL, 154 mmol) and 2,7-dibromofluorene (5.01 g, 15.4 mmol) into a 150 mL one-necked bottle, heat and stir to dissolve. 50 mL of 50% KOH aqueous solution was added to the above reaction system, followed by tetrabutylammonium bromide (0.7 g, 2.17 mmol), and the reaction mixture was heated to 75° C. for 2 h. After the reaction was completed, the reaction mixture was extracted with dichloromethane (50 mL×2), the organic phases were combined, washed with water, 1M hydrochloric acid, washed with water, dried over anhydrous magnesium sulfate, and then the solvent was removed by rotary evaporation. Excess 1,6-dibromohexane was distilled off under reduced pressure to obtain a brown crude product. The crude product was separated and purified by silica gel column chromatography with petroleum ether as the eluent to obtain 2,7-dibromo-9,9-bis(6-bromohexyl)fluorene (7.40 g, 74%). 1 H NMR (300MHz, CDCl 3 ,ppm):7.53(d,2H...

Embodiment 2

[0048] Example 2, PFPN-based FRET technology to detect DNA methylation to be tested

[0049] The following examples select genes whose promoter regions should be unmethylated in normal samples and methylated in cancer samples. According to literature reports, three related genes RASSF1A, OPCML and HOXA9 were selected in the experiment. All gene sequences can be found at http / / :www.ensembl.org. The designed amplified region (belonging to the promoter region) should contain 2 or 3 5'-CCGG-3' sites, where CG methylation is related to ovarian cancer and cannot be at the primer position.

[0050] The acquisition of genomic DNA

[0051] 35 ovarian cancer tissue samples and 11 normal tissue samples were selected from Changzhou Maternal and Child Health Hospital. All samples were obtained with informed consent. Genomic DNA was extracted from formalin-fixed and paraffin-embedded tissue samples with TIANamp gene extraction kit, and the genomic DNA of 35 ovarian cancer tissue samples...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention discloses a reagent kit for detecting cancer-related DNA methylation and application of the reagent kit for detecting cancer-related DNA methylation. The invention provides the application of the substances for detecting CCGG site methylation levels of RASSF1A gene, OPCML gene and HOXA9 gene in the promoter region of the genome of the sample to be detected in the preparation of products for identifying or assisting identification whether the sample to be detected is a cancer sample. Experiments prove that the method and the promoter of the reagent kit for detecting cancer-related DNA methylation are capable of identifying whether the sample to be tested is a cancer sample or whether the subject to be tested is a cancer patient, so that a single sample can be timely and accurately identified; compared with the detection of methylation status of a single gene, the reagent kit for detecting cancer-related DNA methylation has higher diagnostic sensitivity and better specificity; moreover, the statistic analysis and the cumulative analyses are replaced by threshold values, so that instant diagnosis of cancer can be more likely realized.

Description

technical field [0001] The invention relates to a kit and application for detecting cancer-related DNA methylation in the field of biotechnology. Background technique [0002] DNA methylation is an important epigenetic mechanism that plays an important role in the development of human cancer. Aberrant methylation of promoter regions of specific genes has attracted widespread attention as a potential marker for cancer diagnosis. Very few individual genes have been shown to be highly methylated in cancer. Therefore, detecting the methylation status of a group of genes has higher diagnostic sensitivity and specificity than detecting the methylation status of individual genes. The methylation profiles of a variety of cancers have been studied to date. For an ideal methylation profile, accurate detection methods and logical data analysis are essential. In 2012, a method for counting the methylation levels of multiple genes was established, and a step-by-step cumulative analys...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6886C12Q2600/154
Inventor 王树张江艳刘礼兵吕凤婷
Owner INST OF CHEM CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap