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Primer and method for mass spectrometric detection of hotspot mutation of PIK3CA genes by using primer

A technology of hotspots and genes, applied in the field of molecular biology, can solve problems such as low accuracy, low sensitivity, and high cost, and achieve the effects of high accuracy, low detection cost, and high throughput

Active Publication Date: 2013-04-03
武汉康圣达医学检验所有限公司
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Problems solved by technology

[0009] In order to overcome the deficiencies of the above-mentioned prior art, the present invention provides a primer and a method for detecting PIK3CA hotspot mutation by mass spectrometry of the primer; it aims to solve the problems of low sensitivity, low throughput, low accuracy and high cost in the prior art

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  • Primer and method for mass spectrometric detection of hotspot mutation of PIK3CA genes by using primer
  • Primer and method for mass spectrometric detection of hotspot mutation of PIK3CA genes by using primer
  • Primer and method for mass spectrometric detection of hotspot mutation of PIK3CA genes by using primer

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[0047] The technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments. Apparently, the described embodiments are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without creative efforts fall within the protection scope of the present invention.

[0048] The present invention is specifically described below in conjunction with specific samples:

[0049] (1) Determine the position of the PIK3CA hotspot mutation in the gene sequence;

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Abstract

The invention relates to the field of biotechnology, and provides a primer and a method for mass spectrometric detection of hotspot mutation of PIK3CA genes by using the primer. The method comprises the following steps of: (1) positioning hotspot mutation sites of the PIK3CA genes; (2) designing a primer for amplified reaction and a primer for extension reaction according to deoxyribonucleic acid (DNA) sequences at two ends of the mutation sites; (3) performing the amplified reaction; (4) performing treatment by using a serum amyloid protein (SAP) enzyme (alkaline phosphatase); (5) performing the extension reaction; (6) purifying a product of the extension reaction by using resin; (7) transferring the product purified by the resin to a chip; (8) putting the chip in a mass spectrograph, and running a detection procedure; and (9) reading and analyzing detection data, and determining the gene type of a target site of the PIK3CA genes. Compared with the conventional detection method, the method provided by the invention has the characteristics of high sensitivity, high accuracy, high flux and low cost.

Description

technical field [0001] The invention belongs to the field of molecular biology, in particular to a primer and a method for detecting PIK3CA gene hotspot mutation by mass spectrometry of the primer. Background technique [0002] (1), single nucleotide polymorphism [0003] Single nucleotide polymorphism (single nucleotide polymorphism, SNP), mainly refers to the DNA sequence polymorphism caused by the variation of a single nucleotide at the genome level. It is the most common type of heritable variation in humans. Accounting for more than 90% of all known polymorphisms. SNPs exist widely in the human genome, with an average of 1 in every 500-1000 base pairs, and it is estimated that the total number can reach 3 million or more. The polymorphism shown by SNP only involves the variation of a single base, which can be caused by the transition or transversion of a single base, or by the insertion or deletion of a base. Generally speaking, SNP refers to a single nucleotide var...

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 郝玮陈然张聪李金欢李小青周蕊梁超韩韬黄士昂涂赞兵陈忠方国伟
Owner 武汉康圣达医学检验所有限公司
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