Digital PCR method and detection kit for detecting HER2 copy number variation in breast cancer

A detection kit, copy number variation technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve problems such as inability to completely and accurately determine HER2

Active Publication Date: 2019-12-27
NAT INST OF METROLOGY CHINA
View PDF3 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the technical solution of this patent application can only solve the problem of HER2-positive samples that are judged as suspected positive by conventional methods, but cannot solve the samples that are actually false ne...

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
  • Digital PCR method and detection kit for detecting HER2 copy number variation in breast cancer
  • Digital PCR method and detection kit for detecting HER2 copy number variation in breast cancer
  • Digital PCR method and detection kit for detecting HER2 copy number variation in breast cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Example 1 Utilizes ddPCR technology to detect normal human cell DNA and breast cancer cell DNA

[0089] (1) Extraction of cellular genomic DNA:

[0090] Centrifuge the sample, add buffer to suspend, and then add proteinase K to dissolve the sample to obtain a sample solution; add 200 μL of buffer, mix thoroughly by inversion, and place at 70°C for 10 minutes; add 200 μL of absolute ethanol, fully Shake and mix for 15 seconds; add the obtained solution and flocculent precipitate into an adsorption column, centrifuge at 12000rpm (~13400×g) for 30 seconds, discard the waste liquid, put the adsorption column back into the collection tube; add Centrifuge 500 μL of buffer solution at 12000 rpm (~13400×g) for 30 seconds, pour off the waste liquid, put the adsorption column into the collection tube; add 600 μL of washing solution to the adsorption column, centrifuge at 12000 rpm (~13400×g) for 30 seconds, pour Remove the waste liquid, put the adsorption column into the collect...

Embodiment 2

[0103] Example 2 Using ddPCR technology to detect samples with different mutation levels of HER2

[0104] Sample 3 and sample 6 were mixed in different proportions to form 4 samples with different mutation levels of HER2, and the H1 / P1, H2 / P1, and H3 / P1 values ​​of each sample were measured by the method in Example 1. The results are shown in Table 2 below. The ratios of copy number and internal reference gene amplified by the three pairs of primers were all very close to the theoretical values ​​prepared, the correlation coefficients were all greater than 0.93, and the linear correlation coefficients were all greater than 0.995. It shows that the accuracy of the method described in the present invention is high, and the detection results for sample 3 with the lowest mutation level show that the method can accurately detect samples with as little as 1 copy increase, and the determination results of the three pairs of primers are highly consistent.

[0105] Table 2 Determinati...

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 present invention discloses a digital PCR method and a detection kit for detecting HER2 copy number variation in breast cancer. The kit comprises at least one pair of primers and a matching probefor detecting HER2 gene copy number variation and a pair of primers and a matching probe for detecting internal reference gene. The method is conducted by using three sets of HER2 gene specific primerprobes and one set of internal reference gene, thereby effectively eliminating false negative results caused by amplification of a single primer probe. Therefore, compared with prior art, the methodhas higher accuracy, uncertainty interval of the prior art is reduced from (1.8-2.2) to (1.0-1.2), and the method has higher sensitivity.

Description

technical field [0001] The invention relates to the technical field of gene detection related to tumor targeted therapy, in particular to a digital PCR method and a detection kit for detecting HER2 copy number variation of breast cancer. Background technique [0002] Breast cancer is one of the most common malignant tumors in women, with more than 1 million new cases every year. It is a disease that seriously threatens the health and even life of patients. The therapeutic effects and prognosis of clinical drugs may also vary greatly among different patients. [0003] The HER2 gene is a proto-oncogene located on the long arm of human chromosome 17, encoding human epidermal growth factor receptor-2 (human epidermal growth factor receptor-2, HER2) with receptor tyrosine kinase (PTK) activity Membrane glycoprotein, a member of the epidermal growth factor receptor family, needs to combine with other receptors in the family to form a heterodimer to play a signal transduction func...

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): C12Q1/6858
CPCC12Q1/6858C12Q2563/107C12Q2563/159C12Q2545/101C12Q2537/16
Inventor 董莲华王霞王晶
Owner NAT INST OF METROLOGY CHINA
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