Supercharge Your Innovation With Domain-Expert AI Agents!

Method and kit for identifying pulmonary nodule state

A technology of pulmonary nodules and kits, applied in biochemical equipment and methods, microbial determination/testing, DNA/RNA fragments, etc.

Inactive Publication Date: 2021-08-20
BEIJING EXELLON MEDICAL TECH CO LTD
View PDF8 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a lack of means to effectively detect the methylation status of these cancer-related genes and process the detection results

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
  • Method and kit for identifying pulmonary nodule state
  • Method and kit for identifying pulmonary nodule state
  • Method and kit for identifying pulmonary nodule state

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] Example 1: DNA Extraction

[0076] DNA Extraction Reagent consists of Lysis Buffer, Binding Buffer, Washing Buffer and Elution Buffer. Lysis buffer consists of protein denaturants, detergents, pH buffers and nuclease inhibitors. Binding buffers consist of protein denaturants and pH buffers. The cleaning buffer is divided into cleaning buffer A and cleaning buffer B, and cleaning buffer A is composed of protein denaturant, nuclease inhibitor, detergent, pH buffer and ethanol; cleaning buffer B is composed of nuclease inhibitor, pH buffer and ethanol composition. The elution buffer consists of nuclease inhibitors and pH buffers. The protein denaturant is: guanidine hydrochloride; the detergent is: Tween20; the pH buffer is: Tris-HCl; the nuclease inhibitor is: EDTA.

[0077] In this embodiment, the plasma samples of lung cancer patients are taken as an example to extract plasma DNA. The extraction method comprises the following steps:

[0078] (1) Take 1 mL of plasm...

Embodiment 2

[0090] Example 2: Bisulfite Treatment of DNA

[0091] Bisulfite treatment of DNA is to use bisulfite reagent to treat the extracted DNA sample. The bisulfite reagent is composed of bisulfite buffer and protection buffer; the bisulfite buffer is bisulfite A mixture of sodium and water; the protection buffer is a mixture of hydroquinone, an oxygen free radical scavenger, and water.

[0092] In this embodiment, the DNA extracted in Example 1 is used as the processing object, and the DNA is treated with bisulfite, and the specific steps include:

[0093] (1) Preparation of bisulfite buffer: weigh 1g of sodium bisulfite powder, add water to prepare 3M buffer;

[0094] (2) Preparation of protection buffer: weigh 1g of hydroquinone reagent, add water to prepare 0.5M protection buffer;

[0095] (3) Mix 100 μL DNA solution, 200 μl bisulfite buffer solution and 50 μL protection solution, shake and mix evenly;

[0096] (4) Thermal cycle: 95°C for 5 minutes, 80°C for 60 minutes, 4°C fo...

Embodiment 3

[0107] Example 3: miRNA extraction, reverse transcription

[0108] The miRNA extraction reagent consists of lysis buffer, binding buffer, washing buffer and elution buffer. Lysis buffer consists of protein denaturants, detergents, pH buffers and nuclease inhibitors. Binding buffers consist of protein denaturants and pH buffers. The cleaning buffer is divided into cleaning buffer A and cleaning buffer B, and cleaning buffer A is composed of protein denaturant, nuclease inhibitor, detergent, pH buffer and ethanol; cleaning buffer B is composed of nuclease inhibitor, pH buffer and ethanol composition. The elution buffer consists of nuclease inhibitors and pH buffers. Wherein the protein denaturant is: guanidine hydrochloride and 2-ME; the detergent is: Tween20; the pH buffer is: Tris-HCl; the nuclease inhibitor is: EDTA.

[0109] In this embodiment, plasma samples of patients with lung cancer were taken as an example to extract plasma miRNA. The extraction method comprises t...

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 provides a method for identifying a pulmonary nodule state in a subject. The method comprises the following steps of: 1) detecting the methylation level of biomarker genes and / or the expression level of miRNA in a biological sample from the subject, wherein the biomarker genes comprise PTGER4, RASSF1A and SHOX2 genes, and the miRNA comprises miR-19b and miR-320a; and 2) comparing the methylation level detected in the step 1) with the normal methylation level of corresponding biomarkers in a population, and / or comparing the expression level of the miRNA detected in the step 1) with the normal expression level of the corresponding miRNA so as to identify the pulmonary nodule state of the subject. The invention also provides a kit for identifying the pulmonary nodule state in the subject. The method and the kit provided by the invention provide a rapid, reliable and accurate new way for identification of benign and malignant pulmonary nodules and prediction, diagnosis and evaluation of lung cancer.

Description

technical field [0001] This article relates to methods and kits for identifying the status of pulmonary nodules in subjects, especially methods and kits for identifying the status of pulmonary nodules by using the methylation levels of biomarker genes and / or the expression levels of miRNAs. Background technique [0002] Lung cancer is a malignant tumor with the highest morbidity and mortality rate in the world, and it is a major disease that threatens human life and health. In 2014, the World Cancer Report released by the International Agency for Research on Cancer (IARC) of the World Health Organization showed that the global cancer burden is increasing, and lung cancer ranks first among the new common cancers in 2012, accounting for about 1.8 million cases, accounting for 20% of the total number of common cancers. 13%; in the common death cancer, lung cancer also ranks first, about 1.6 million cases, accounting for 19.4% of the total. Among the top ten cancers with high i...

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): C12Q1/6883C12N15/113
CPCC12Q1/6883C12Q2600/178C12Q2600/158C12Q2600/154
Inventor 徐春叶李明明蒲珏
Owner BEIJING EXELLON MEDICAL TECH CO LTD
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com