Specific primes and liquid phase chip for AGT (angiotensinogen) and ACE (angiotensin-converting enzyme) gene polymorphism detection

Abstract

The invention discloses specific primes and a liquid phase chip for AGT (angiotensinogen) and ACE (angiotensin-converting enzyme) gene polymorphism detection. The liquid phase chip mainly contains ASPE (allele specific primer extension) which consists of tag sequences at 5' end and specific primers at 3'-end relative to a target gene mutation, wherein the specific primers respectively are SEQ ID NO. 9 and SEQ ID NO. 10, SEQ ID NO. 11 and SEQ ID NO. 12, SEQ ID NO. 13 and SEQ ID NO. 14 regarding to relative to the AGT gene, and / or SEQ ID NO. 15 and SEQ ID NO. 16 relative to the ACE gene; microspheres coated with anti-tag sequences; and amplification primers. The conformation between the detection result of the AGT and ACE gene polymorphism detection liquid phase chip and the result of a sequencing method is up to 100%. The prepared liquid phrase has excellent signal-noise ratio and can avoid cross reaction and realize the parallel detection of multiple polymorphic sites.

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 AGT and ACE gene polymorphism and a liquid phase chip. Background technique [0002] Renin-angiotensin-aldosterone (renin-angiotensin system, RAS) is a hormone system that plays an important role in the regulation of blood pressure, blood flow and internal environment. Among them, angiotensinogen and angiotensin converting enzyme It is a key component of the RAS system. [0003] Angiotensinogen (Angiotensinogen, AGT) is an α-2 globulin that is continuously synthesized by the liver and released into the blood circulation. The main function of AGT is to play an important role in regulating blood pressure as a substrate of renin. Angiotensin-Converting Enzyme (ACE) is also known as dipeptide carboxypeptidase. The main function of ACE is to convert angiotensin I into angiotensin II, which has a strong vasoconstri...

AI Technical Summary

Problems solved by technology

Fluorescent quantitative PCR has the advantages of simple operation, quick results, and quantification. However, this technology has the disadvantages of easy sample contamination, cross-reaction, and high false positive rate; while PCR-RFLP method is based on restriction endonucleation caused by gene mutations. Changes in the enzyme recognition site, such as site loss or new site generation, a specific fragment is amplified by PCR, and then the amplified product is digested with a restriction endonuclease, and the size of the fragment is observed by electrophoresis. This method is used for Detection of gene mutations with altered restriction sites can directly determine the genotype, but 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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