VHL (Von Hippel Lindau) genetic mutation detection specific primer and liquid phase chip

Abstract

The invention discloses a VHL (Von Hippel Lindau) genetic mutation detection specific primer and a liquid phase chip. The liquid phase chip mainly comprises ASPE (Allele Specific Primer Extension) primers, microspheres coated by an anti-tag sequence, and amplimers, wherein the ASPE primers consist of tag sequences at a 5' end and specific primer sequences specific to target genetic mutation sites at a 3' end; and the specific primer sequences are: SEQ ID NO.13 and SEQ ID NO.14 specific to a T240A site; SEQ ID NO.15 and SEQ ID NO.16 specific to a T254C site; SEQ ID NO.17 and SEQ ID NO.18 specific to a T266A site; SEQ ID NO.19 and SEQ ID NO.20 specific to a T286T site; SEQ ID NO.21 and SEQ ID NO.22 specific to a G388C site; and / or SEQ ID NO.23 and SEQ ID NO.24 specific to a 444delT site. The matching rate between a detection result of the liquid phase chip provided by the invention and a sequencing method is up to 100 percent, and wild and mutant parallel detection of a plurality of mutant sites is realized.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a VHL gene mutation detection specific primer and a liquid phase chip. Background technique [0002] VHL gene is named after Von Hippel-Lindau (VHL) disease, contains 14543bp, includes 3 exons and 2 introns. VHL disease is a clinically very rare familial autosomal dominant genetic tumor disease, including central nervous system hemangioblastoma, visceral tumor and cyst, etc. In 1993, Latif et al. located the VHL gene on chromosome 3p25-26 through linkage analysis , and successfully cloned the VHL gene for the first time. The VHL gene is a tumor suppressor gene. The inactivation of this gene will lead to the disorder of normal VHL protein synthesis, which is closely related to the occurrence and development of clear cell renal cell carcinoma (CCRCC). At present, important progress has been made in the research on the structure and function ...

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.

Images