CYP2C8 gene polymorphism detection specific primers and liquid chip

Abstract

The invention discloses CYP2C8 gene detection specific primers and a liquid chip. The liquid chip mainly comprises: various ASPE primers composed of 5' terminal tag sequence and 3' terminal specific primer sequences aiming at target gene mutation site, microspheres coated with anti-tag sequence, and amplification primers. The specific primer sequences comprise: SEQ ID NO.13 and SEQ ID NO.14 aiming at A127T site; SEQ ID NO.15 and SEQ ID NO.16 aiming at A124G site; SEQ ID NO.17 and SEQ ID NO.18 aiming at G145A site; SEQ ID NO.19 and SEQ ID NO.20 aiming at 204delA site; SEQ ID NO.21 and SEQ ID NO.22 aiming at C115G site; and / or SEQ ID NO.23 and SEQ ID NO.24 aiming at G315C site. The matching rate of the detection result obtained by using the liquid chip provided by the invention and a result obtained by using a sequencing method is 100%. Therefore, wild-type and mutant-type parallel detections of a plurality of mutation sites are realized.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a CYP2C8 gene polymorphism detection specific primer and a liquid phase chip. Background technique [0002] The cytochrome P450 enzyme system is a superfamily consisting of enzyme proteins encoded by homologous genes. There are 57 CYP genes in the human genome, which encode corresponding CYP enzymes, among which the CYP1, CYP2 and CYP3 families are the most important. CYP2C8 is an important hepatic CYP enzyme, which accounts for 7% of the total liver CYP and participates in nearly 15% of drug catalysis in phase I metabolism. The anticancer drug paclitaxel is the specific metabolic substrate of CYP2C8, and the 6a-hydroxyl CYP2C8 is the most commonly used marker reaction for in vitro detection of CYP2C8, and CYP2C8 is also the main CYP enzyme that converts arachidonic acid into active eicosatrienoic acid in liver and kidney. [0003] The CYP2...

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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