Kit and method for detecting gene polymorphism based on shared primer probes and application of method or kit

Abstract

The present application discloses a method and kit for detecting gene polymorphism based on shared primer probes and an application of the method or the kit. According to the method, reporter groups are labeled at the 5' ends of a wild-type ARMS primer and a mutant ARMS primer codirectionally designed, and ARMS primers with probe performance are prepared and used for gene polymorphism detection after being screened, especially for polymorphism detection of human CYP2C19. The method is simple in design, low in cost, and has higher detection sensitivity, stronger specificity, and more accurate and objective results.

Description

[0001] This divisional application is based on the division of the Chinese patent application with the application number 201911055308.5, the application date is October 31, 2019, and the invention title is "a kit, method and application for detecting genetic polymorphisms based on shared primer probes". case application. technical field [0002] This application relates to the field of gene detection, in particular to a PCR detection of gene polymorphism. [0003] technical background [0004] 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 w...

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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