Kit, method and application for detecting gene polymorphism based on shared primer probe

Abstract

The present application discloses a method, kit and application for detecting gene polymorphism based on shared primer probes. In the method, the reporter group is labeled on the 5' ends of the wild-type ARMS primers and mutant ARMS primers designed in the same direction to prepare the probe-specific ARMS primers, which are used for gene polymorphism detection after screening, especially for Human CYP2C19 polymorphism detection. The method has simple design, low cost, higher detection sensitivity, stronger specificity, and more accurate and objective results.

Description

technical field [0001] This application relates to the field of gene detection, in particular to a PCR detection of gene polymorphism. [0002] technical background [0003] At present, the methods used to detect gene polymorphism mainly include the following: sequencing typing method, high resolution melting curve typing method, gene chip typing method, TaqMan probe typing method and MGB probe typing method, etc. . Sequencing typing is the gold standard for detection in this field, but it has not been widely promoted due to the cumbersome operation and special equipment. The other methods are derived from PCR amplification technology and are relatively widely used. Among them, the detection process of sequencing typing method includes PCR amplification and sequencing, and the operation is cumbersome. The detection time is generally as long as 4 to 5 hours, and the PCR reaction tube needs to be opened during the operation process, which is easy to cause contamination of PCR ...

AI Technical Summary

Problems solved by technology

Sequencing typing is the gold standard for detection in this field, but it has not been widely promoted due to the cumbersome operation and special equipment; the rest of the methods are derived from PCR amplification technology and are relatively widely used

Among them, the detection process of sequencing typing method includes PCR amplification and sequencing, and the operation is cumbersome. The detection time is generally as long as 4 to 5 hours, and the PCR reaction tube needs to be opened during the operation process, which is easy to cause contamination of PCR products; high resolution The characteristic of the melting curve typing method is to use the difference in Tm value caused by the difference of the base of the polymorphic site, and detect by identifying different Tm values. 1°C, so equipment that can accurately identify the temperature is needed. Such equipment is relatively rare, and most of them are imported and expensive. The detection process of gene chip typing method includes PCR amplification and probe hybridization. The operation is cumbersome, and the detection time is generally as long as 4 hours. ~5 hours, and the PCR reaction tube needs to be opened during the operation, which is likely to cause contamination of PCR products; the TaqMan probe typing method uses ARMS primers to identify polymorphic sites, and this method detects a polymorphic site Two reaction wells are required, and the throughput is relatively low; MGB probe typing uses MGB-labeled probes to identify polymorphic sites, MGB probe design is difficult and expensive, and finding a suitable MGB probe requires screening A lot of lines, the cost is higher

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