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Allele variant detection method, kit and composition

An allele and detection method technology, applied in the direction of recombinant DNA technology, DNA/RNA fragments, etc., can solve the problem of low detection rate, and achieve the effect of improving sensitivity and specificity, and improving amplification efficiency.

Inactive Publication Date: 2013-07-24
SHENZHEN UNI MEDICA TECH
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  • Abstract
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Problems solved by technology

[0005] The object of the present invention is to provide a kind of allelic variant detection method, kit and method for the low detection rate of allelic variant detection for existing probe hybridization method such as AS-PCR or genotyping combination

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  • Allele variant detection method, kit and composition
  • Allele variant detection method, kit and composition
  • Allele variant detection method, kit and composition

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Embodiment Construction

[0051] The present invention mainly relates to detection methods, kits and compositions of allelic variants. The main terms involved have the following definitions:

[0052] Allelic variant: A variant form of one of two or more genes that occurs at a particular locus on a chromosome, including single nucleotide polymorphisms (SNPs), insertions, inversions, Bits and missing.

[0053] Primer: An oligonucleotide that hybridizes to a strand of nucleotides in a target sequence.

[0054]The alleles detected in the present invention are derived from accounting samples, and the accounting samples can use any nucleic acid molecule, which can be DNA, such as genomic DNA (gDNA) or complementary DNA (cDNA), or RNA, such as messenger RNA ( mRNA), etc.

[0055] The allelic variant detection method of the present invention specifically comprises the following steps:

[0056] 1) Add mutant allele-specific primers and their reverse primers and a blocker whose 3' end cannot be extended by p...

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Abstract

The invention provides a method for carrying out allele variant specific detection by utilizing an inhibitor, a kit and a composition. The method provided by the invention comprises the following steps of: adding a primer pair and an inhibitor the terminal 3' of which can not be extended by polymerase in a hybridization process into a nucleic acid sample, wherein the primer pair comprises a specific primer and a reverse primer; hybridizing the specific primer and the reverse primer respectively with target sequence mutant type oligonucleotide in the nucleic acid sample to produce an amplicon; hybridizing the inhibitor with a target sequence wild type oligonucleotide in the nucleic acid sample; and carrying out real-time specific detection on a mutant type allele by utilizing a fluorescent dye or a detection pointer. The added inhibitor can not extend in an amplification process, and amplification of a wild type allele variant is inhibited, so that the amplification efficiency of a mutant type allele variant is improved, and the sensitivity and specificity of detection are improved.

Description

technical field [0001] The invention relates to the field of gene detection, more specifically, to a method kit and composition for detecting allelic variants by using allele-specific polymerase chain reaction (AS-PCR). Background technique [0002] Single nucleotide polymorphism (SNP) is the most common type of genetic diversity in the human genome, and its frequency in human genomic DNA is about 1 SNP in 1,000 nucleotides, or less. SNPs have been implicated in genetic disorders, susceptibility to different diseases, predisposition to adverse reactions to drugs, and are used in forensic research. Therefore, the detection of SNPs (or rare mutations) provides great potential for early diagnosis of diseases, such as detection of circulating cancer cells in blood for prenatal diagnosis, and detection of disease-associated mutations in mixed cell populations, The use of SNPs to identify target genes related to diseases or drug responses is the focus of current research. [000...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
Inventor 罗晓腾
Owner SHENZHEN UNI MEDICA TECH
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