Unlock instant, AI-driven research and patent intelligence for your innovation.

Universal reagent for nucleic acid detection

A reagent and molecular detection technology, applied in the field of DNA hybridization reaction, can solve problems such as difficult to use in field environment without detection conditions, complex equipment and detection system, etc.

Pending Publication Date: 2022-06-28
BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods can realize the quantification of the substances to be tested, but they also require complex equipment and detection systems, which are difficult to use in the field environment without detection conditions

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
  • Universal reagent for nucleic acid detection
  • Universal reagent for nucleic acid detection
  • Universal reagent for nucleic acid detection

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1. Oligonucleotide strands required for CHA to catalyze the hairpin assembly reaction.

[0055] The oligonucleotide chain of CHA reaction includes C, H, F chain, and its complementary structure is such as figure 2 shown (SEQ ID NOs: 1-4). The C chain firstly hybridizes with the neck loop of the H chain through the TMSD reaction, and opens the hairpin structure of H, and then the H chain is complementary to the neck loop of the F chain through the TMSD reaction, and opens the hairpin structure of F to form the H-F-C complex. Finally, the free end of the F chain replaces C through the TMSD reaction, so that C is separated from the hybrid complex to form an H-F complex, and C acts as a catalyst to participate in another assembly process. In order to facilitate the TMSD reaction of the H and F chains immobilized on different microspheres, the 5'-end (or all 3'-ends) of the H and F chains are biotin (Biotin) or amino (-NH) 2 ) group is combined with the correspond...

Embodiment 2

[0056] Example 2. Conjugation of oligonucleotide probe strands to microspheres

[0057] The H and F chains can be tightly linked to avidin microspheres by labeling biotin at the 5' or 3'-end, or labeling with amino groups (-NH 2 ) and the carboxyl functional group (-COOH) on the surface of the carboxyl microspheres through chemical cross-linking reaction to form chemical bonds. The cross-linking between the amino group and the carboxyl group adopts the classical method, using carbodiimide (EDC) and N-hydroxysuccinimide (NHS) as the cross-linking agent. The principle is as follows: EDC first reacts with a carboxyl group to form an O-acylisourea intermediate that can react with amino groups, and NHS is used to stabilize the intermediate. After the addition of amino-modified terminal nucleotides, this intermediate can rapidly react with an amino group to form an amide bond, covalently labeling the oligonucleotide chain to polystyrene microspheres.

[0058] The end of the F chai...

Embodiment 3

[0061] Example 3. Comparison of agglutination reactions of different microspheres

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 the technical field of nucleic acid molecule detection, in particular to a probe combination and application thereof. According to the present invention, the toehold-mediated strand displacement (TMSD, Toehold-mediated strand displacement) reaction is performed between the nucleic acid chains connected with the microspheres, such that the microspheres form the agglutination blocks so as to be visible to the naked eyes; the method does not need complex operation or expensive equipment, does not need biological enzyme participation, does not need to amplify a target to be detected, and can be used as a molecular detection reagent to provide rapid and on-site qualitative and semi-quantitative detection. Experiments show that the probe combination designed by the invention can accurately identify samples containing targets and samples without targets in the process of detecting the samples, and the sensitivity can reach 100fM.

Description

technical field [0001] The present invention relates to the technical field of nucleic acid molecule detection, in particular to DNA hybridization reaction and application thereof. Background technique [0002] Molecular diagnosis has a wide range of applications in the health field, and can be used in clinical diagnosis, infectious disease prevention and control, and environmental pollution detection. Nucleic acid diagnosis in molecular diagnosis, that is, to detect whether a nucleic acid of a specific sequence is contained in the test object, is the most widely used. Nucleic acid diagnostic markers of various specific sequences are an important basis for disease diagnosis. Existing diagnostic methods mainly rely on the amplification of nucleic acid fragments mediated by enzymes, such as PCR, and temperature amplification reactions such as LAMP, NASBA, etc., which are often limited by low-temperature storage, transportation, and the cost of enzyme preparation. It is a nuc...

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): C12N15/11C12Q1/682C12Q1/6834
CPCC12Q1/682C12Q1/6834C12Q2525/301C12Q2537/1373C12Q2563/149
Inventor 康禹邵长君王建楚亚男陈婧
Owner BEIJING INST OF GENOMICS CHINESE ACAD OF SCI CHINA NAT CENT FOR BIOINFORMATION