ABCC (ATP (adenosine triphosphate)-binding cassette, sub-family C)1 gene mutation detection specific primers and liquid chip

Abstract

The invention discloses an ABCC (ATP (adenosine triphosphate)-binding cassette, sub-family C)1 gene mutation detection liquid chip and specific primers. The liquid chip mainly comprises each ASPE (allele specific primer extension) primer formed by tag sequences at 5' terminal and specific primer sequences aiming at target gene mutation sites at 3' terminal, microspheres coated by anti-tag sequences as well as amplification primers, wherein the specific primer sequences are SEQ ID NO.15 and SEQ ID NO.16 aiming at A866T sites, SEQ ID NO.17 and SEQ ID NO.18 aiming at C218T sites, SEQ ID NO.19 and SEQ ID NO.20 aiming at G2168A sites, SEQ ID NO.21 and SEQ ID NO.22 aiming at G3173A sites, SEQ ID NO.23 and SEQ ID NO.24 aiming at G16237754A sites, SEQ ID NO.25 and SEQ ID NO.26 aiming at C16238494T sites and / or SEQ ID NO.27 and SEQ ID NO.28 aiming at G1299T sites. The liquid chip has the advantages that the coincidence rate of the detection results of the detection liquid chip provided by the invention and a sequencing method is as high as 100%; and 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 ABCC1 gene mutation detection and a liquid phase chip. Background technique [0002] The ABCC subfamily belongs to the ABC transporter superfamily, and ABCC1 is the first discovered member of the ABCC subfamily. ABCC1 is also known as multidrug resistance-related protein 1 (mul tidrug resistance protein 1, MRP1), located on chromosome 16 16p13.1, its encoded protein is distributed in almost all tissues in the body, and can transport a variety of different substrates, including drugs , heavy metal ions, body metabolic poisons, glutathione, glucuronic acid and sulfur conjugates. With the development of sequencing technology, many polymorphisms of ABCC1 gene have been found in recent years, and more and more evidences show that ABCC1 gene variation is closely related to drug resistance and disease susceptibility. [0003...

AI Technical Summary

Problems solved by technology

[0003] At present, ABCC1 gene mutation detection methods mainly include: PCR-RFLP, direct sequencing method and fluorescent quantitative PCR technology. PCR-RFLP method is based on the change of restriction enzyme recognition site caused by gene mutation, such as site loss or generation New site, a specific fragment is amplified by PCR, and then the amplified product is digested with restriction endonuclease, and the size of the fragment is observed by electrophoresis. Genotype, but this method cannot be used for the detection of gene mutations that do not generate new restriction sites

However, other PCR-based detection technologies, such as direct sequencing and fluorescent quantitative PCR technology, have the disadvantages of low sensitivity, easy contamination of samples, and high false positive rate.

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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