Point mutation gene detection method and kit thereof

A mutation gene and point mutation technology, which is applied in the field of a method and kit for detecting point mutation genes, can solve the problems of low detection sensitivity of gene point mutations and wild-type gene interference, and achieve high detection sensitivity and accurate detection results , the effect of good application prospects

Inactive Publication Date: 2013-07-10
SHANGHAI BIOCHIP
View PDF0 Cites 9 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention aims to solve the technical problems that the current point mutation detection sensitivity is not high, and the detection process is easily interfered by wild-type genes

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
  • Point mutation gene detection method and kit thereof
  • Point mutation gene detection method and kit thereof
  • Point mutation gene detection method and kit thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1 Detection of point mutation genes by the method of the present invention

[0036] 1. Design two specific probes

[0037] For the common mutation site Y220C of the p53 gene, two specific probes for the mutation base are designed, and their structures are as follows:

[0038]

[0039] Among them, the last base C at the 3' end of probe I (SEQ ID NO.1) is just in the complementary base pairing where the point mutation occurs, and an oligo that has nothing to do with the target sequence is labeled at the 5' end of this probe. Nucleotide sequence (italic part) is used as the recognition sequence, which is used in the subsequent reaction to pair and hybridize with the complementary sequence on the surface of the magnetic microsphere, thereby isolating the connected probe; probe II (SEQ ID NO.2) and the mutant gene After complementary pairing, it is adjacent to the previous probe, and the 5' end is adjacent to the base where the point mutation occurs, and the 3' e...

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 point mutation gene detection method. The method comprises the following steps: forming a complete oligonucleotide chain by treating a mutant gene as a template and connecting two oligonucleotide probes complementary and adjacent to the point mutation gene through a connection reaction, and marking the oligonucleotide chain with electrochemical luminescence molecules; and capturing through magnetic microspheres to separate out the complete oligonucleotide chain, detecting the electrochemical luminescence molecules marked on the complete oligonucleotide chain through using an electrochemical luminescence analyzer, and comparing the electrochemical luminescence signal intensities of a gene sample to be tested and a blank solution to find the difference in order to detect the point mutation gene. The invention also discloses a pint mutation gene detection kit. The kit includes two oligonucleotide probes complementary to the point mutation gene, a ligase, and magnetic microspheres having surfaces marked with the capturing molecules. The point mutation gene detection method has the advantages of high detection sensitivity, and accurate and reliable detection result, and has a very good application prospect in the tumor molecule diagnosis field, the genetic molecule diagnosis field and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method and a kit for detecting point mutation genes. Background technique [0002] With the development of molecular biology, more and more evidences show that major diseases such as malignant tumors and genetic diseases are closely related to gene mutations, and point mutations are the most common types of gene mutations. Mutation gene detection technology has been developed rapidly. At present, gene sequencing technology is the "gold standard" for detecting gene mutations. However, due to the interference of wild-type gene background when detecting mutant genes, sequencing technology is limited by sensitivity, and it is difficult to detect mutant genes with an abundance of less than 10%. Also due to the interference of wild-type genes, polymerase chain reaction (PCR) cannot amplify and enrich low-abundance mutant genes, and it is difficult to increase the proportion ...

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): C12Q1/68G01N21/76
Inventor 程昌明汪杰杨超
Owner SHANGHAI BIOCHIP
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