Two-part mediator probe

An intermediary and probe technology, applied in the fields of biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of non-universal methods, a lot of work and time, and achieve the effect of simplifying primer design

Pending Publication Date: 2019-12-03
ALBERT LUDWIGS UNIV FREIBURG +1
View PDF9 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Assays described in the literature that allow the simultaneous detection of several molecular classes are either not universal (Das et al., 2012) or have the additional disadvantage that the detection reaction must be carried out in several stages (Linardy et al., 2016), which Requires a lot of work and time during implementation

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
  • Two-part mediator probe
  • Two-part mediator probe
  • Two-part mediator probe

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0244] Example 1: Mediation Probe

[0245] Embodiments of the inventive design include an intermediary probe for detecting at least one target molecule, wherein the intermediary probe comprises at least two oligonucleotides. The first oligonucleotide has a Mediator binding region and a probe region. The Mediator binding region is located at the 5' end of the oligonucleotide and the probe region is located at the 3' end of the oligonucleotide. A second or several other oligonucleotides, a mediator or mediators, that chemically, biologically and / or physically bind to the mediator binding region of the first oligonucleotide . Mediators may consist of DNA, RNA, PNA or modified RNA such as LNA. The probe region of the first oligonucleotide has an affinity for the target molecule and / or the template molecule, and the mediator binding region has an affinity for a mediator or a plurality of mediators ( figure 1 ). A mediator or a plurality of mediators has an affinity for at lea...

Embodiment 2

[0246] Embodiment 2: the program of mediator replacement

[0247] After binding of the probe region to the target molecule and / or the template molecule, for example using a strand displacement polymerase, the Mediator is displaced by the Mediator binding region. This process can occur during the amplification of target molecules and / or template molecules. In an embodiment of the present invention, the probe region of the intermediary probe can be used as a primer in DNA amplification. After binding of the probe region to the target molecule and / or the template molecule, the intermediary probe is extended. A second primer can then be attached to the extended mediation probe and extended. During the amplification process, a mediator (mediator) or multiple mediators (mediators) are released from the mediator-binding region and trigger a detectable signal by interacting with one or more detection molecules ( figure 2 ).

Embodiment 3

[0248] Example 3: Detection Molecule with 6 Regions

[0249] The released, unlabeled mediator is detected with the help of a detection reaction. The reaction mechanism described below can be carried out in parallel with the amplification of target molecules and / or template molecules as described above.

[0250] In a preferred form of the invention, the detection molecule may consist of an oligonucleotide divided into six regions ( image 3 ). Region 1 includes the 5' end of the detection molecule consisting of the sequence segment and the fluorescent acceptor Q. Region 3 is the reverse complement of region 1 and is separated by region 2. Region 4 separates region 3 from region 5, and region 4 can specifically interact with Mediator molecules. Region 6 comprises a sequence region at the 3' end which may have chemical modifications and thus allow directional immobilization of oligonucleotides. Fluorescent donors F are associated in a suitable manner with the regions from re...

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 relates to a mediator probe for detecting at least one target molecule, comprising at least two oligonucleotides. A first oligonucleotide of the mediator probe according to the inventioncomprises a probe region and a mediator binding region, wherein the probe region has an affinity for a target molecule and / or template molecule, and the mediator binding region has an affinity for atleast one mediator. At least one additional oligonucleotide of the mediator probe is a mediator which is bound to the first oligonucleotide of the mediator probe via the mediator binding region and has an affinity for at least one detection molecule. The mediator triggers a detectable signal after the first oligonucleotide of the mediator probe is released by interacting with the detection molecule. The invention additionally relates to a system comprising at least one mediator probe according to the invention and at least one detection module and to a method for detecting at least one targetmolecule.

Description

[0001] introduction [0002] The present invention relates to a mediation probe for detecting at least one target molecule comprising at least two oligonucleotides. The first oligonucleotide of the Mediator probe according to the present invention comprises a probe region and a Mediator binding region, wherein the probe region has an affinity for the target molecule and / or the template molecule, and the Mediator binding region has an affinity for at least one Mediator The body has affinity. The at least one additional oligonucleotide of the Mediator probe is a Mediator, which binds to the first oligonucleotide of the Mediator probe via a Mediator binding region and has an affinity for at least one detection molecule, wherein, in the After release of the first oligonucleotide of the probe, the mediator triggers a detectable signal through interaction with the detection molecule. Furthermore, the invention relates to a system comprising at least one mediator probe according to t...

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(China)
IPC IPC(8): C12Q1/6844C12Q1/6823
CPCC12Q1/6823C12Q1/6844C12Q2525/161C12Q2525/205C12Q2525/301C12Q2527/101C12Q2537/137C12Q2561/101C12Q1/6832C12Q1/6853C12Q1/686
Inventor 马丁·特罗特西蒙·瓦德勒费利克斯·冯·施特滕莉萨·贝赫勒
Owner ALBERT LUDWIGS UNIV FREIBURG
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