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

A kind of fish probe set and application thereof for detecting ntrk fusion

A probe group and probe technology, applied in the direction of DNA/RNA fragments, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of continuous activation of kinase region, abnormal expression of TRK, low expression level, etc. , improve the quality of life, the effect of high probe specificity and sensitivity

Active Publication Date: 2022-06-07
AMOYDX BIOTECHNOLOGY RES CENT CO LTD
View PDF7 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The expression level of TRK in normal tissues is low, but when fusion occurs (the partner gene is fused with the kinase region at the 3' end of NTRK as the 5' end), it can lead to abnormal expression of TRK, resulting in continuous activation of the kinase region and causing tumors

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
  • A kind of fish probe set and application thereof for detecting ntrk fusion
  • A kind of fish probe set and application thereof for detecting ntrk fusion
  • A kind of fish probe set and application thereof for detecting ntrk fusion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] like figure 1 As shown, the NTRK1 5′-end probe set (the centromeric side of the NTRK1 gene) used two BAC clones (RP11-1054F3 and RP11-66D17) as templates, and the NTRK1 3′-end probe set (the telomere side of the NTRK1 gene) ) using two BAC clones (RP11-1038N13 and RP11-110J1) as templates; NTRK2 5′-end probe (the centromere side of NTRK2 gene) using two BAC clones (RP11-324L15 and CTD-2509D1) as templates, Two BAC clones (RP11-152D13 and CTD-2339E11) were used as templates for NTRK2 3′ end probe (telomere side of NTRK2 gene); 2 BAC clones were used for NTRK3 5′ end probe (telomere side of NTRK3 gene) (RP11-110O23 and RP11-97O12) were used as templates, and two BAC clones (RP11-113C11 and CTD-2526N1) were used as templates for NTRK3 3'-end probes (the centromeric side of NTRK3 gene). The 5'-end probes of the three genes were all labeled with FITC (the first fluorescent reporter group) as a green signal, and the 3'-end probes were all labeled with Tetramethyl-rhodamine (...

Embodiment 2

[0062] The steps for determining the labeling efficiency and concentration of the probes in the above probe set using an ultraviolet spectrophotometer are as follows:

[0063] (1) Turn on the UV spectrophotometer (NanoDrop), select the full wavelength mode, and select the corresponding wavelength. After blank calibration, measure the A2 of the 5' end probe group respectively 60nm , A Dye (FITC: A 494nm ) and A of the 3' end probe set 260nm , A Dye (Tetramethyl-rhodamine: A 560nm ).

[0064] (2) Since the fluorescent dye also has a certain absorbance value at 260nm, in order to make the absorbance value of the probe at 260nm more accurate, use the following formula to correct A 260 value: A Base =A 260 -(A Dye ×CF 260 ), CF 260 refers to the correction factor, which is a constant and has a defined CF for each fluorochrome 260 value, as CF of Tetramethyl-rhodamine 260 The value is 0.06 and the FITC is 0.32. (3) Calculate the probe labeling efficiency and concentrati...

Embodiment 3

[0069] The NTRK1 gene fragmentation probe system and NTRK3 gene fragmentation probe system prepared in Example 1 were used to detect 50 samples of papillary thyroid cancer provided by the hospital, and the NTRK3 gene fragmentation probe system was used to detect 10 samples of salivary gland mammary gland secretory carcinoma, NTRK2 The gene fragmentation probe system was used to detect 20 pontine glioma samples, and the NTRK probe from Empire Genomics was used to detect these samples. The detection steps are as follows:

[0070] (1) Deparaffinization and rehydration of the sample: immerse the tissue section samples in 3 cylinders of xylene for dewaxing for 10 minutes each time, and then soak them in 100%, 100%, 100%, 85% and 70% ethanol for 3 minutes. A final immersion in deionized water for 3 min.

[0071] (2) Sample pretreatment and digestion: boil the tissue slice samples in water at 100°C for 20 minutes, take out the samples and digest them with pepsin for 10 minutes, trans...

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 FISH probe set for detecting NTRK fusion and its application, including NTRK1 5' end probe set, NTRK1 3' end probe set, NTRK2 5' end probe set, NTRK2 3' end probe set , NTRK3 5' end probe set and NTRK3 3' end probe set. The invention is quick and easy to operate, with accurate and reliable detection results, can accurately detect all fusion types of NTRK1, NTRK2, and NTRK3 genes, provides a reference for the treatment plan of tumor patients, is beneficial to individualized targeted therapy of patients, reduces drug side effects, and improves patient Quality of life, reduce family and social burden, suitable for large-scale promotion and application.

Description

technical field [0001] The invention belongs to the technical field of molecular detection, in particular to a FISH probe set for detecting NTRK fusion and its application. Background technique [0002] NTRK1, NTRK2, and NTRK3 belong to the neurotrophic receptor tyrosine kinase (NTRK) family, located on chromosomes 1, 9, and 15, respectively, and are responsible for encoding the tropomyosin receptor kinase (TRK) family proteins TrkA, TrkB, and TrkC, respectively. When the extracellular ligand neurotrophic factor binds to the Trk protein receptor, it can induce the receptor to form a dimer and undergo phosphorylation, activate RAS / MAPK / ERK, P13K, PLCγ and other downstream signaling pathways, and participate in cell growth, proliferation, etc. process. The expression level of TRK in normal tissues is low, but when fusion occurs (partner gene as 5' end fused with NTRK 3' end kinase domain), it can lead to abnormal expression of TRK, resulting in continuous activation of the ki...

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 Patents(China)
IPC IPC(8): C12Q1/6886C12Q1/6841C12N15/11
CPCC12Q1/6886C12Q1/6841
Inventor 卢皇彬林煌艺朱思琪郑立谋
Owner AMOYDX BIOTECHNOLOGY RES CENT CO LTD
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