ABCB2 gene polymorphism detection specific primers and liquid chip

Abstract

The invention discloses ABCB2 gene detection specific primers and a liquid chip. The liquid chip mainly comprises: various ASPE primers composed of tag sequence and specific primer sequences aiming at 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 G142A site; SEQ ID NO.15 and SEQ ID NO.16 aiming at G88A site; SEQ ID NO.17 and SEQ ID NO.18 aiming at G154A site; SEQ ID NO.19 and SEQ ID NO.20 aiming at C86T site; SEQ ID NO.21 and SEQ ID NO.22 aiming at C150T site; and / or SEQ ID NO.23 and SEQ ID NO.24 aiming at A92T 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 specific primer for detecting ABCC2 gene polymorphism and a liquid phase chip. Background technique [0002] Multidrug resistance-related protein ABCC2 (MRP2) is widely distributed in the small intestine, liver and kidney of mammals, and is specifically located on the apical membrane of polar cells. At present, there are many methods for detecting and analyzing the polymorphism of the ABCC2 gene, such as: direct sequencing, Real time-PCR, PCR-RFLP analysis, etc., among which the most commonly used method is PCR-RFLP analysis. 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. The size of the fragment is observed b...

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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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