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

Liquid chip for detecting gene point mutation and method

A liquid phase chip, point mutation technology, applied in biochemical equipment and methods, microbial determination/inspection, fluorescence/phosphorescence, etc., can solve the problems of low sensitivity of gene point mutation detection, wild-type gene interference, etc. Accurate and reliable results, enhanced signal-to-noise ratio, and improved sensitivity

Active Publication Date: 2013-05-15
SHANGHAI BIOCHIP
View PDF0 Cites 5 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 detection sensitivity of gene point mutations is not high, and the detection process is easily interfered by wild-type genes in samples, and provides a liquid phase chip for detecting gene point mutations, which is suitable for a large number of wild-type gene backgrounds Detection of low-abundance point mutation 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
  • Liquid chip for detecting gene point mutation and method
  • Liquid chip for detecting gene point mutation and method
  • Liquid chip for detecting gene point mutation and method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Detection of samples by the liquid phase chip of the present invention

[0047] 1. A pair of specially designed specific primers

[0048] A pair of specific primers were designed for the common mutation site Y220C of the P53 gene, the structure of which is as follows:

[0049]

[0050] The forward primer is coated on the surface of the microsphere, and the first base C at the 3' end is complementary to the base of the mutation site of the gene to be detected, and is not complementary to the wild-type gene; the third base A at the 3' end is An artificially introduced base that is not complementary to the mutant and wild-type genes; the 10 base A at the 5' end is the connecting arm between the primer sequence and the microsphere. The 5' end of the reverse primer was modified with biotin as a signal reporter molecule.

[0051] 2. Amplification of mutant genes in water-in-oil emulsion

[0052] (1) The volume of the aqueous phase is 72 μl, and the amplification...

Embodiment 2

[0068] Example 2 Detection of samples by the liquid phase chip of the present invention

[0069] 1. A pair of specially designed specific primers

[0070] A pair of specific primers were designed for the common mutation site Y220C of the P53 gene, the structure of which is as follows:

[0071]

[0072] The forward primer is coated on the surface of the microsphere, and the first base C at the 3' end is complementary to the base of the mutation site of the gene to be detected, and is not complementary to the wild-type gene; the third base A at the 3' end is An artificially introduced base that is not complementary to the mutant and wild-type genes; the 6 carbon atoms (C6) at the 5' end are the connecting arms between the primer sequence and the microsphere. The 5' end of the reverse primer was modified with CY5 fluorescein as a signal reporter.

[0073] 2. Amplification of mutant genes in water-in-oil emulsion

[0074] (1) The volume of the aqueous phase is 100 μl, and th...

Embodiment 3

[0088] Embodiment 3 Comparison experiment between the present invention and allele-specific primer extension method

[0089] Allele-specific primer extension (Allele specific primer extension, ASPE) is a common method for detecting gene point mutations. For the primer, the base that is different from the normal allele is arranged at the 3' end of the primer, so that the primer is completely paired with the mutant gene, and an extension reaction occurs after the primer binds to the mutant gene template; while the 3' end of the primer The terminal base is a mismatch with the wild-type gene, and no extension reaction occurs after combining with the wild-type gene template, and whether the sample contains a mutant gene can be detected by detecting the extension product.

[0090] This embodiment compares the sensitivity of the present invention and the ASPE method in detecting gene point mutations. In order to facilitate visual comparison with the detection results of the present ...

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
The average particle sizeaaaaaaaaaa
Login to View More

Abstract

The invention discloses a liquid chip for detecting gene point mutation. The liquid chip comprises: a primer pair designed specific to gene mutation sites, a microsphere, and a signal report molecule. In the primer pair, the last basic group at the 3' terminal of a forward primer is complementary to the basic group of a to-be-detected gene mutation site, and is not complementary to a wild-type gene. A basic group that is not complementary to mutant-type and wild-type genes is introduced into a forward primer sequence artificially. The forward primer is in connection with the microsphere through a connecting arm. A reverse primer is completely complementary to the mutant-type gene, and its 5' terminal is labeled with the signal report molecule. The invention also discloses a method for detecting gene point mutation. The method comprises the steps of: designing the primer pair specific to gene mutation sites; using the primer pair to conduct amplification on a to-be-detected mutation gene in a water-in-oil emulsion; and detecting a fluorescence signal on the microsphere surface. With the liquid chip and the method for detecting gene point mutation provided in the invention, the detection sensitivity of low abundance gene point mutation under the background of a lot of wild-type genes can be improved substantially.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a liquid phase chip and a method for detecting gene point mutations. Background technique [0002] With the progress of the Human Genome Project, more and more evidences show that gene mutations play an important role in human diseases. Through the detection of gene mutations, diseases can be effectively prevented, diagnosed and treated. Gene mutation refers to the change of gene structure caused by the substitution, addition or deletion of DNA base pairs, which includes point mutations caused by single base changes, or multiple base deletions, duplications and insertions, in which point Mutations are most common. [0003] The physical and chemical properties of genes caused by point mutations change very little, which undoubtedly makes the detection of point mutations very difficult. Especially when the abundance of the mutated gene to be detected is low and exists in ...

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): C12Q1/68G01N21/64
Inventor 程昌明
Owner SHANGHAI BIOCHIP
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