High-sensitivity mutation site detection system, high-sensitivity mutation site detection method and application

A detection method and system technology, applied in the field of biomedicine, can solve the problem of limited ability to detect rare gene mutations

Active Publication Date: 2018-10-12
SUZHOU VERSABIO TECH INC
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Problems solved by technology

However, in the context of higher wild-type templates, the ability to detect rare gene mutations is limited

Method used

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  • High-sensitivity mutation site detection system, high-sensitivity mutation site detection method and application
  • High-sensitivity mutation site detection system, high-sensitivity mutation site detection method and application
  • High-sensitivity mutation site detection system, high-sensitivity mutation site detection method and application

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Embodiment Construction

[0070] The present invention will be further described in detail below in conjunction with the embodiments, so that those skilled in the art can implement it with reference to the description.

[0071] It should be understood that terms such as "having", "comprising" and "including" used herein do not exclude the presence or addition of one or more other elements or combinations thereof.

[0072] A highly sensitive gene mutation detection method (ASPB) of this embodiment includes: designing wild-type upstream or downstream primers for wild-type genes, mutant upstream or downstream primers for mutant genes, and universal upstream or downstream primers , mutant blocker and wild type blocker;

[0073] A double-tube PCR system is used for PCR reaction, and quantitative PCR detection is performed on samples; one tube includes wild-type primers, mutant blocker and universal primers, and the other tube includes mutant primers, wild-type blocker and universal primers.

[0074] Wherei...

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Abstract

The invention discloses a high-sensitivity mutation site detection system, a high-sensitivity mutation site detection method and application. The high-sensitivity mutation site detection method comprises the following steps of designing wild-type forward or reverse primers aiming at wild-type genes, mutant forward or reverse primers aiming at mutant genes, universal forward or reverse primers, mutant blockers and wild-type blockers; using a double-tube PCR system for PCR reaction; and performing quantitative PCR detection on a sample, wherein one tube comprises wild-type primers, mutant blockers and universal primers, and the other tube comprises mutant primers, wild blocks and universal primers. The high-sensitivity mutation site detection method provided by the invention has the advantages that by using a method of combining specific primers with specific blockers, the specificity and the sensitivity of the detection are improved. Through the standard product and standard curve drawing, the false positive and false negative results are well avoided. The capability of detecting gene mutation of ten thousandth or higher can be realized.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a highly sensitive gene mutation detection system, method and application. Background technique [0002] Gene mutation refers to changes in the composition or sequence of base pairs in the structure of genes, such as single nucleotide polymorphisms (SNPs) and changes in bases after unmethylated nucleotides are converted. [0003] Single nucleotide polymorphisms (SNPs) refer to the polymorphisms caused by the substitution of a single nucleotide (A, G, C, T) in the genomic DNA sequence, and are a new generation of polymorphic genetic markers. SNPs widely exist in biological genomes, for example, there are more than 3 million SNPs in the entire genome, for example, there are 3 billion bases in human beings every 1,000 bases. SNPs are closely related to the occurrence and development of diseases. [0004] The methylation of cell genes is closely related to the occurrence and development ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6858
CPCC12Q1/6858C12Q2531/113C12Q2525/186
Inventor 刘兆成赵国栋
Owner SUZHOU VERSABIO TECH INC
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