Liquid phase chip for CYP19A1 gene SNP (Single Nucleotide Polymorphism) detection and detection method thereof

Abstract

The invention discloses a liquid phase chip for CYP19A1 gene SNP (Single Nucleotide Polymorphism) detection, which comprises wild type and mutant type specific ASPE primers designed targeting CYP19A1 gene SNPs of rs4646, rs10046C>T, rs700519C>T, rs1870050C>A, hCV1664178A>C, rs12900137G>C, rs730154G>A, rs936306T>C and rs1902586A>G, microballoon spheres respectively coated with specific anti-tag sequences and primers used for amplifying CYP19A1 gene SNPs with target sequences to be detected. Each ASPE primer comprises a tag sequence at 5' end and a specific primer sequence at 3' end, wherein the specific primers are selected from SEQ ID NO. 19-36 and the tag sequences are selected from SEQ ID NO.1-18; and the anti-tag sequences can be correspondingly in complementary pairing with the tag sequences. The invention also discloses a CYP19A1 gene SNP detection method. The coincidence rate of the detection method provided by the invention and a sequencing method is as high as 100%; and the prepared liquid phase chip for CYP19A1 gene SNP (Single Nucleotide Polymorphism) detection has excellent signal-to-noise rate, and basically no cross reaction exists between a design probe and the anti-tag sequences.

Description

technical field [0001] The invention belongs to the field of molecular biology and relates to medicine and biotechnology, in particular to a CYP19A1 gene SNP detection liquid chip and a detection method. Background technique [0002] In recent years, aromatase inhibitors (Aromatase inhibitors, AIs) have received more and more attention and application in the treatment of breast and ovarian cancer due to their strong specificity, high efficacy, and small side effects. Aromatase belongs to the cytochrome P450 family, which can catalyze the conversion of androgen into estrogen. It is the rate-limiting enzyme in the synthesis of estrogen. closely related to the occurrence of cancer. Therefore, AIs can reduce the level of estrogen by inhibiting aromatase, thereby eliminating the stimulating effect of estrogen on tumor growth. Both in vivo and in vitro studies have shown that the third-generation AIs drugs, such as letrozole and anastrozole, can effectively inhibit the transform...

AI Technical Summary

Problems solved by technology

[0006] At present, there are products on the market to detect CYP19A1 gene polymorphisms, such as Applied Biosystems' SNPGenotyping Assays series products are allelic difference analysis methods based on TaqMan technology, but they can only detect one mutation at a time, which is time-consuming and laborious; the traditional solid-phase chip method is expensive and has low sensitivity. poor repeatability

However, other PCR-based techniques for detecting SNPs, such as direct sequencing, semi-quantitative PCR, and PCR-single-strand conformational polymorphism (SSCP) detection, have low sensitivity, easy sample contamination, and high false positive rates. Due to the limitations of detection throughput, ordinary PCR methods and fluorescent quantitative PCR cannot meet the needs of detection and promotion.

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