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

Method for determining a mutation in genomic dna, use of the method and kit for carrying out said method

A genome and gene technology, applied in the field of implementing the method or kit for the purpose, can solve the problems of unnecessary treatment and wrong treatment of patients

Active Publication Date: 2018-06-08
迪莫迪特里希
View PDF10 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But on the other hand, it is also possible that patients get unnecessary treatment or receive it at the wrong time

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 for determining a mutation in genomic dna, use of the method and kit for carrying out said method
  • Method for determining a mutation in genomic dna, use of the method and kit for carrying out said method
  • Method for determining a mutation in genomic dna, use of the method and kit for carrying out said method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 9

[0197] In a preferred use of the method for prediction, a prediction of a patient's response to treatment with at least one active ingredient selected from the group consisting of alkylating chemotherapeutics, cytostatics, therapeutic monoclonal antibodies and inhibitory agents, especially tyrosine kinase inhibitors. Particular preference is given to making predictions regarding active ingredients selected from the group consisting of Temozolomide, Cetuximab, Bevacizumab, AG-221, AGI-5198, AG-120, AG-881 or combinations thereof.

[0198] In a preferred variant of the method for the follow-up of a patient's malignant disease it is contemplated that steps B) and C) are in a manner that allows quantitative determination of the methylation status of at least one mutation and / or at least one CpG-dinucleotide under conditions. Preferably, the use of the method for follow-up of a malignant disease then comprises the steps of: i) providing a first sample with genomic DNA from a patie...

Embodiment 1

[0216] Example 1: Determination of point mutations in untransformed genomic DNA (reference)

[0217] Point mutations are those in which only a single nucleobase is affected by the change. The reliable determination of point mutations therefore places high demands on the specificity and sensitivity of molecular diagnostic assays.

[0218] As an example for a point mutation of high clinical relevance, the point mutation V600E within the BRAF gene was studied.

[0219] Genomic DNA to be analyzed can come from a variety of sources. For example, fixed tissue is used here. Three thin sections of 10 μm each of formalin-fixed and paraffin-embedded malignant melanoma and one tissue block of normal tissue (skin) adjacent to the melanoma were transferred to separate 2 ml reaction vessels. Subsequently, genomic DNA was extracted from the tissue sections. For this purpose the use of the QIAamp DNA FFPE tissue kit (Qiagen, Hilden, Germany) according to the manufacturer's instructions ...

Embodiment 2

[0223] Example 2: Determining Point Mutations in Genomic DNA

[0224] In the extracted DNA from Example 1, in each case part of the genomic DNA was transformed according to the invention. Conversion can be accomplished, for example, by contacting the genomic DNA with bisulfite. Conversion is currently performed eg with the innuCONVERT bisulfite all-in-one kit (Analytik Jena, Jena, Germany). For this purpose, 2 μg of extracted DNA from each sample was transformed according to the manufacturer's instructions. The amount of converted DNA was then quantified using a NanoDrop ND-1000 spectrophotometer (Thermo Fisher Scientific, Waltham, MA, USA).

[0225] In the normal state, DNA is organized in the form of a double helix, which consists of two single strands that are reverse complementary to each other. One strand is called the positive or forward strand and the other is called the negative or reverse strand. After DNA conversion, eg with bisulfite, the plus and minus strand...

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

Disclosed is a method for determining a mutation in genomic DNA. The method is characterised in that the mutation analysis is carried out using genomic DNA in which at least one portion of the cytosine contained therein has previously been converted into uracil or into another base with a base pair behaviour or molecular weight that differs from cytosine.

Description

[0001] Citations from earlier applications [0002] This patent application claims priority from German Patent Application No. 10 2015 009 187.5, the disclosure content of which is incorporated herein by reference. [0003] sequence listing [0004] This application includes as part of the specification an electronic sequence listing of 94 sequences in txt format according to the WIPO ST.25 standard. field of invention [0005] The present invention relates to molecular diagnostic methods in the field of oncology for the determination of mutations in genomic DNA. The invention further relates to the use of such molecular diagnostic methods in relation to the diagnosis, prognosis, prediction and follow-up of malignant diseases. Furthermore the invention relates to kits for carrying out said method or for said use. In particular the method according to the invention is an in vitro method. Background technique [0006] Targeted therapy plays an important role in the era of ...

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): C12Q1/70C12Q1/6827C12Q1/6886
CPCC12Q1/6827C12Q1/6886C12Q1/708C12Q2600/154C12Q2600/156C12Q2523/125C12Q1/6806
Inventor 迪莫·迪特里希
Owner 迪莫迪特里希
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