Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Blood circulation miRNA biomarker assay kit for diagnosis and prognosis evaluation of gastric cancer

A technology of biomarkers and detection kits, applied in the field of kits, can solve the problems of insufficient protein stability, improper storage, easy degradation, and insurmountability, and achieve the effect of excellent diagnostic efficiency and good application prospects

Pending Publication Date: 2019-09-13
内蒙古医科大学附属人民医院
View PDF0 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In summary, the existing problems in the prior art are: the diagnostic specificity and sensitivity of traditional tumor markers CEA, CA199, and CA724 in gastric cancer are not high
CEA is expressed as a protein. Due to its own biological characteristics, the protein is not stable enough, and it is easy to degrade if stored improperly. This is also the reason for the low specificity and sensitivity of this type of tumor marker, and its disadvantages cannot be overcome in clinical testing.

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
  • Blood circulation miRNA biomarker assay kit for diagnosis and prognosis evaluation of gastric cancer
  • Blood circulation miRNA biomarker assay kit for diagnosis and prognosis evaluation of gastric cancer
  • Blood circulation miRNA biomarker assay kit for diagnosis and prognosis evaluation of gastric cancer

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0095] (1) Preparation of 2.5% agarose gel plate

[0096] Add 2.5g of agarose powder to 0.5×TBE100ml and heat it in a microwave oven until the powdered agarose is completely dissolved; when cooled to 60°C-70°C, add 10mg / ml ethidium bromide 5ul, pour into the sealed gel tank , with a thickness of about 3-5mm, insert the toothed comb; after cooling and forming, carefully pull out the toothed comb, put it into the agarose horizontal electrophoresis tank, and submerge the gel surface with 0.5xTBE until 1cm;

[0097] (2) Take 0.3 μg of total RNA, add 1 / 5 volume of 5× loading buffer, heat at 65°C for 5 minutes, and quench on ice to eliminate the secondary structure of RNA; add 0.5 μg to the RNA sample before loading -1.0μl ethidium bromide EtBr, concentration 1.0mg / mL; prepared 1.2% formaldehyde denaturing gel in 1×formaldehyde denaturing gel electrophoresis buffer for pre-electrophoresis for 15min; RNA samples at a voltage of 5-10V / cm Lower the electrophoresis for 30min.

[0098]...

Embodiment 1

[0104] Gastric cancer detection kit

[0105] (1) Nucleotide sequence mir-30c, specific miR-30c nucleotide sequence:

[0106] 5'-TGTAAACATCCTACACTCTCAGC-3'.

[0107] (2) U6 snRNA is used as an internal reference: the sequences of primers upstream and downstream of U6 primers are:

[0108] 5'-CTCGCTTCGGCAGCACA-3',

[0109] 5'-AACGCTTCACGAATTTGCGT-3'.

[0110] (3) Calculation method of relative expression of miR-30c: 2 -ΔCt Cycle value calculation, ΔCt=Ct miR-30c -Ct U6

[0111] 1. Instrument

[0112] The main instruments are as follows

[0113]

[0114]

[0115] 2. Kit detection steps

[0116] 1. Kit detection:

[0117] 1.1 Treatment of serum samples All the included populations took 3ml of fasting venous blood in the morning, and immediately centrifuged with a horizontal centrifuge at 2000rpm for 8min to separate the serum. The supernatant was placed in a new centrifuge tube and stored in a -80°C refrigerator for extraction of total RNA. Extraction of total RN...

Embodiment 2

[0163] 2.1 Sample collection:

[0164] Conduct clinical sample research in Inner Mongolia Autonomous Region, China. A total of 240 inpatient gastric cancer surgery samples were collected from the First Affiliated Hospital of Inner Mongolia Medical College from January 2006 to October 2011. The removed gastric cancer tissue samples were immediately frozen in liquid nitrogen and then stored at -80°C in the refrigerator. The above surgical resection samples were independently diagnosed as gastric cancer by two senior pathologists, and pathological data were collected after pathological examination. The depth of tumor invasion is classified according to UICC (International Union Against Cancer standard) [19] , the status of lymph node metastasis, and the degree of differentiation were judged according to WHO (World Health Organization) standards [20] , The location of the tumor is subject to the pathological report.

[0165] Non-cancerous samples matched by sex, age and gastri...

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 belongs to the technical field of kits, and discloses a blood circulation miRNA biomarker assay kit for diagnosis and prognosis evaluation of gastric cancer. The assay method with the blood circulation miRNA biomarker assay kit for diagnosis and prognosis evaluation of gastric cancer comprises steps of processing serum samples, blood collection, centrifugation, storage for later use;extracting total RNA, storing the extracted total RNA in a refrigerator at 80 DEG C for later use; conducting absorbance detection, determining the quality of the RNA by RNA formaldehyde degradationgel electrophoresis; obtaining cDNA by reverse transcription; conducting fluorescence quantitative detection for the obtained cDNA; and analyzing the detected results by relative quantitative method.Through evaluation of efficacy in diagnosis compared with traditional tumor markers CEA, CA199 and CA724, the blood circulation miRNA biomarker assay kit of the invention is superior to the traditional tumor markers. The miR-30c is suitable for detection of gastric cancer as a diagnostic marker, and can be developed into a novel tumor detection marker diagnosis kit for diagnosis of gastric cancer.The blood circulation miRNA biomarker assay kit is used for evaluation of curative effect and prognosis estimation, and has a good application prospect.

Description

technical field [0001] The invention belongs to the technical field of kits, and in particular relates to a detection kit for gastric cancer diagnosis and prognosis evaluation blood circulation miRNA biomarkers. Background technique [0002] At present, the existing technologies commonly used in the industry are as follows: [0003] Gastric cancer is a common malignant tumor of the digestive system, and its morbidity and mortality are high in my country. According to statistical analysis, the incidence rate of gastric cancer in my country in 2012 was 31.28 / 100,000, and the death rate was 22.04 / 100,000. Gastric cancer often has no obvious specific clinical manifestations and poor prognosis, and the five-year survival rate is only 20-30%. Therefore, it is of great significance to find serum specific markers for gastric cancer for early diagnosis and treatment of gastric cancer. The diagnostic specificity and sensitivity of traditional tumor markers CEA, CA199, CA724 in gastr...

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
IPC IPC(8): C12Q1/6886
CPCC12Q1/6886C12Q2600/158C12Q2600/178C12Q2600/118C12Q2600/166
Inventor 牟永平卫星王振飞贺晓花侯婧田晓燕米澜陈永霞
Owner 内蒙古医科大学附属人民医院
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com