Specific primer and liquid chip for detecting polymorphism of SLC22A1 gene

Abstract

The invention discloses a liquid chip and specific primer for detecting an SLC22A1 gene. The liquid chip mainly comprises ASPE (allele specific primer extension) primers, microspheres and amplification primers, wherein each ASPE primer is formed by tag sequences at the 5' terminal and specific primer sequences which are arranged at the 3' terminal and aim at the mutation site of the target gene; the specific primer sequences are SEQ ID NO.19 and SEQ ID NO.20 aiming at the C117T site, SEQ ID NO.21 and SEQ ID NO.22 aiming at the T198C site, SEQ ID NO.23 and SEQ ID NO.24 aiming at the C86T site, SEQ ID NO.25 and SEQ ID NO.26 aiming at the C97G site, SEQ ID NO.27 and SEQ ID NO.28 aiming at the C164T site, SEQ ID NO.29 and SEQ ID NO.30 aiming at the G123T site, SEQ ID NO.31 and SEQ ID NO.32 aiming at the G127A site, SEQ ID NO.33 and SEQ ID NO.34 aiming at the A148G site and / or SEQ ID NO.35 and SEQ ID NO.36 aiming at the G129C site; and the microspheres are enveloped by anti-tag sequences. The detecting liquid chip provided by the invention has the advantages that the consistency of the detection result and the sequencing method is as high as 100%, thus parallel detection of wild types and mutant types of a plurality of mutation sites is achieved.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a specific primer for detecting SLC22A1 gene polymorphism and a liquid phase chip. Background technique [0002] Organic cation transporter 1 (OCT1, encoding gene SLC22A1) is a transporter with genetic polymorphisms, mainly distributed in the human liver, and mediates the transport of a series of organic cations such as some drugs, poisons and endogenous compounds. SLC22A1 plays an important role in the absorption, distribution and excretion of many exogenous drugs. Studies have shown that the polymorphism of the SLC22A1 gene is the main reason for the individual differences in its transport function. [0003] At present, there are few methods for detecting and analyzing the polymorphism of SLC22A1 gene, mainly including direct sequencing and PCR-RFLP analysis, among which the most commonly used method is PCR-RFLP analysis. The PCR-RFLP me...

AI Technical Summary

Problems solved by technology

The PCR-RFLP method is based on the change of the restriction endonuclease recognition site caused by gene mutation, such as the loss of the site or the generation of a new site, a specific fragment is amplified by PCR, and then cut and amplified with a restriction endonuclease. Amplify the product and observe the size of the fragment by electrophoresis. This method is used to detect gene mutations with altered restriction sites, and can directly determine the genotype. However, this method cannot be used for the detection of gene mutations without new restriction sites.

Again, the above methods all have limitations in detection throughput, and can only detect one mutation type at a time, which cannot meet the needs of practical applications.

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