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

Method for Detecting a Mutation in a Microsatellite Sequence

a microsatellite sequence and mutation technology, applied in the field of microsatellite sequence mutation detection, can solve the problems of inability to efficiently or reliably hybridize the probe, the use of such a technique has never been envisioned for the detection of a mutated microsatellite sequence, and the inability to discriminate between wt and mutant microsatellite sequences, etc., to achieve the effect of increasing sensitivity

Inactive Publication Date: 2020-09-17
INSTITUT CURIE +2
View PDF0 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent describes a method called digital PCR (ddPCR) that allows for very precise measurement of small differences in DNA copy numbers between samples. This method uses a process called partitioning to separate and count each molecule of DNA separately, which makes it easy to measure the abundance of targets in a sample without interference from other molecules. This also allows for the measurement of ratios between targets, which can be used to calculate absolute ratios between them. Overall, this method provides a more accurate and reliable way to measure small changes in DNA copy numbers.

Problems solved by technology

However, the use of such a technique has never been envisioned for the detection of a mutated microsatellite sequence.
Indeed, due to the size and more particularly to the extreme repetitive nature of the microsatellite sequence, it would be expected that the probe covering the microsatellite (MS probe, see below) would slip over the repeat sequence such that efficient or reliable hybridization of the probe could not be obtained.
The Light Cycler HybProbes hybridization probes used in this document are not capable of discriminating WT and mutant microsatellite sequences.
Besides, the probes of Dietmaier et al. are not regarded as hydrolysis probes and are not relevant in the context of a digital PCR reaction.

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 Detecting a Mutation in a Microsatellite Sequence
  • Method for Detecting a Mutation in a Microsatellite Sequence
  • Method for Detecting a Mutation in a Microsatellite Sequence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0035]A—Definitions

[0036]The following definitions are intended to assist in providing a clear and consistent understanding of the scope and detail of the following terms, as used to describe and define the present invention:

[0037]As used herein, the verb “comprise” as is used in this description and in the claims and its conjugations are used in its non-limiting sense to mean that items following the word are included, but items not specifically mentioned are not excluded. In addition, reference to an element by the indefinite article “a” or “an” does not exclude the possibility that more than one of the elements are present, unless the context clearly requires that there is one and only one of the elements. The indefinite article “a” or “an” thus usually means “at least one.”

[0038]As used herein a “tumor” or a “neoplasm” (both terms can be used interchangeably) is an abnormal new growth of cells. The cells in a neoplasm usually grow more rapidly than normal cells and will continue...

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

PropertyMeasurementUnit
fluorescenceaaaaaaaaaa
fluorescence intensityaaaaaaaaaa
sizeaaaaaaaaaa
Login to View More

Abstract

The invention relates to a method for detecting a mutation in a microsatellite sequence locus of a target fragment from a DNA sample, comprising subjecting said DNA sample to a digital polymerase chain reaction (PCR) in the presence of a PCR solution comprising:a pair of primers for amplifying said target fragment of the DNA sample including said microsatellite sequence;a first MS oligonucleotide (MS) hydrolysis probe, labeled with a first fluorophore, wherein said first MS oligonucleotide probe is complementary to a wild-type sequence including the microsatellite sequence;a second oligonucleotide reference (REF) hydrolysis probe, labeled with a second fluorophore, wherein said second oligonucleotide REF probe is complementary to a wild-type sequence of said target DNA fragment which does not include said microsatellite sequence. The invention also encompasses methods for the diagnosis and prognosis of cancer and a method for determining the efficacy of a cancer treatment.

Description

SEQUENCE LISTING SUBMISSION VIA EFS-WEB[0001]A computer readable text file, entitled “SequenceListing.txt,” created on or about Jan. 6, 2020 with a file size of about 4 kb contains the sequence listing for this application and is hereby incorporated by reference in its entirety.INTRODUCTION[0002]Microsatellites (MS) are tandem repeats of short DNA sequences that are abundant throughout the human genome. Owing to their high mutation rates, microsatellite sequences have been widely used as polymorphic markers in population genetics and forensics. Microsatellite instability (MSI) is a hypermutator phenotype that occurs in tumors with impaired DNA mismatch repair (MMR) and is characterized by widespread length polymorphisms of microsatellite repeats due to DNA polymerase slippage as well as by elevated frequency of single-nucleotide variants (SNVs). MSI is caused by inactivation of MMR genes (for example, MLH1, MSH2, MSH3, MSH6 and PMS2) through somatic mutations, with increased risk 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
Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/6858C12Q1/6886G01N21/64
CPCG01N21/6428C12Q1/6858G01N2021/6439C12Q1/6886C12Q2600/156C12Q2525/151C12Q2535/131C12Q2561/101C12Q2563/159
Inventor PROUDHON, CHARLOTTEKASPEREK, AMÉLIEBORTOLINI SILVEIRA, AMANDABIDARD, FRANÇOIS-CLÉMENTSTERN, MARC-HENRI
Owner INSTITUT CURIE
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