Specific primers and liquid phase chip for polymorphic detection of age-related maculopathy susceptibility 2 (ARMS2) gene and high temperature factor A-1 (HTRA1) gene

Abstract

The invention relates to a liquid phase chip for the polymorphic detection of an age-related maculopathy susceptibility 2 (ARMS2) gene and a high temperature factor A-1 (HTRA1) gene. The liquid phase chip comprises allele specific primer extension (ASPE) primers consisting of tag sequences at the 5' end and specific primers aiming at a mutant target gene at the 3' end, microspheres which are wrapped with anti-tag sequences and an amplification primer, wherein the specific primers are SEQ ID No. 10 and SEQ ID No. 11 for a G72T mutant locus of the ARMS2 gene, SEQ ID No. 12 to SEQ ID No. 14 for a C95G / A mutant locus of the HTRA1 gene, SEQ ID No. 15 and SEQ ID No. 16 for a G60A mutant locus of the HTRA1 gene and / or SEQ ID No. 17 and SEQ ID No. 18 for the insertion / deletion (I / D) mutation of the HTRA1 gene. The coincidence rate of a detection result of the liquid phase chip and a sequencing method is up to 100 percent, cross reaction can be prevented, and the parallel detection of multiplepolymorphic loci is 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 ARMS2 and HTRA1 gene polymorphisms and a liquid phase chip. Background technique [0002] Age-related macular degeneration (AMD) is a common eye aging disease caused by degenerative changes in the macula, which can gradually destroy central vision and eventually lead to blindness. AMD with soft drusen, pigmentary abnormalities, and geographic atrophy is often referred to as dry AMD, whereas AMD with choroidal neovascularization, retinal pigment epithelial (RPE) detachment, or discoid fibrosis is termed wet or neoplastic Vascular AMD. In recent decades, AMD has become one of the main causes of blindness in western developed countries. With the aging population in my country, the number of retinal diseases such as AMD is also increasing, and its pathogenesis and treatment methods are receiving more and more att...

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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