Nucleic acid composition, kit and detection method for detecting methylation of lung cancer related genes

A nucleic acid composition and methylation technology, applied in the field of molecular biology, can solve the problem of lack of relevant reagents or kits for simultaneous detection, and achieve the effects of avoiding uneven baselines, flat baselines, and high amplification efficiency

Pending Publication Date: 2020-03-27
SUREXAM BIO TECH
View PDF14 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, there is currently a lack of related reagents or ki

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
  • Nucleic acid composition, kit and detection method for detecting methylation of lung cancer related genes
  • Nucleic acid composition, kit and detection method for detecting methylation of lung cancer related genes
  • Nucleic acid composition, kit and detection method for detecting methylation of lung cancer related genes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] The characteristics and performance of the present invention will be described in further detail below in conjunction with the examples.

[0083] This embodiment provides a kit for detecting the methylation of lung cancer-related genes, the lung cancer-related genes RASSF1A, SHOX2 and PTGER4 genes, the kit includes: primer probe mixture, hot start Taq DNA polymerase, 2× methylation Kylation detection PCR buffer.

[0084]Wherein, the primer-probe mixture includes: a primer-probe combination for detecting RASSF1A, SHOX2 and PTGER4 gene methylation and an ACTB internal reference gene primer-probe combination, the concentration of each primer is 1.25 μM, and the concentration of each probe is 0.5 μM. The sequences of each primer and probe are shown in Table 1 below.

[0085] Table 1

[0086]

[0087]

[0088] In Table 1, the 5' end of the RASSF1A gene methylation detection probe is labeled with the fluorescent reporter group FAM, and the 3' end is labeled with the q...

Embodiment 2

[0121] Embodiment 2 alveolar lavage fluid sample detection

[0122] Select 50 cases of alveolar lavage fluid samples from patients with known lung cancer and patients with benign lung diseases (pneumonia, tuberculosis, etc.), take 15 mL of alveolar lavage fluid, centrifuge at 1000×g for 5 minutes to collect cells, use commercial kits to extract cell DNA, and Using the detection method provided by the present invention to detect the methylation of RASSF1A, SHOX2 and PTGER4 genes on DNA samples. The detection results of alveolar lavage fluid samples from patients with lung cancer and patients with benign lung diseases are shown in Table 5-6. According to the above detection data, it can be seen that the sensitivity of lung cancer detection is 84% ​​and the specificity is 94% when the kit of the present invention is used to detect the samples of alveolar lavage fluid.

[0123] Table 5 Ct values ​​detected in alveolar lavage fluid samples of patients with lung cancer

[0124] ...

Embodiment 3

[0130] Example 3 Plasma sample detection

[0131] Select known lung cancer patients and benign lung disease (pneumonia, pulmonary tuberculosis, etc.) plasma samples of 50 cases each, get 4mL of plasma and use a commercial kit to extract plasma free DNA, and use the detection method provided by the present invention to carry out RASSF1A, SHOX2 and PTGER4 gene methylation detection. The test results of alveolar lavage fluid samples from patients with lung cancer and patients with benign lung diseases are shown in Table 7-8. According to the above detection data, it can be seen that the sensitivity of lung cancer detection is 64% and the specificity is 96% when using the kit of the present invention to detect plasma samples.

[0132] Table 7 Ct values ​​detected in plasma samples of patients with lung cancer

[0133]

[0134]

[0135] Table 8 Ct values ​​detected in plasma samples of patients with benign lung disease

[0136]

[0137]

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 nucleic acid composition, kit and detection method for detecting methylation of lung cancer related genes, and relates to the field of molecular biology. The nucleic acid composition comprises a primer composition, wherein the primer composition comprises a first primer pair for detecting methylation of an RASSF1A gene, a second primer pair for detecting methylation of anSHOX2 gene and a third primer pair for detecting methylation of a PTGER4 gene. The nucleic acid composition can realize simultaneous detection of methylation of the RASSF1A, HOX2 and PTER4 genes on the basis of a fluorescent quantitative PCR platform, and has the characteristics of high sensitivity and high specificity.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, and in particular relates to a nucleic acid composition, a kit and a detection method for detecting the methylation of lung cancer-related genes. Background technique [0002] There are about 781,000 cases of lung cancer in my country each year, and about 626,000 cases of death. It is currently the cancer with the highest morbidity and mortality in my country. Due to the lack of obvious symptoms of early lung cancer and the insensitivity of the existing early diagnosis methods, most lung cancers are discovered at an advanced stage, and the treatment methods are limited. The 5-year survival rate is only 15.8%. "Chinese Expert Consensus on Diagnosis and Treatment of Pulmonary Nodules (2018 Edition)" pointed out that the prevalence of pulmonary nodules in China is nearly 100 million, of which 5-10% are lung cancer. If lung cancer can be detected at this stage a...

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): C12Q1/6886C12Q1/686C12N15/11
CPCC12Q1/6886C12Q1/686C12Q2600/154C12Q2600/16C12Q2563/107C12Q2545/114
Inventor 曾杰彭璨璨吴诗扬刘志明
Owner SUREXAM BIO TECH
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