THADA gene mutation detection specific primer and liquid phase chip

Abstract

The invention discloses a THADA gene mutation detection liquid phase chip and a specific primer. The liquid phase chip mainly comprises: an ASPE primer composed of a tag sequence at 5'-terminal and specific primer sequences against the mutation site of a target gene at 3'-terminal, wherein the specific primer sequences comprise SEQ ID NO.13 and SEQ ID NO.14 against an A109G site, SEQ ID NO.15 and SEQ ID NO.16 against a T156C site, SEQ ID NO.17 and SEQ ID NO.18 against an A89C site, SEQ ID NO.19 and SEQ ID NO.20 against a G153A site, SEQ ID NO.21 and SEQ ID NO.22 against a G162T site, and/or SEQ ID NO.23 and SEQ ID NO.24 against a C80T site; a microsphere coated with an anti-tag sequence; and an amplimer. The coincidence rate between the detection result of the detection liquid phase chip and a sequencing method is 100%, and the wild and mutant parallel detection of a plurality of mutation sites can be realized.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a THADA gene mutation detection specific primer and a liquid phase chip. Background technique [0002] The THADA gene is a thyroid adenoma-associated gene, and its English name is thyroid adenoma associated, which encodes a thyroid adenoma-related protein. The THADA gene is located on the short arm of chromosome 2 at 2p21, has 38 exons, is about 365kb in length, and is widely expressed in various tissues throughout the body, especially the liver. Their meta-analysis of genome-wide association studies found that the THADA gene was associated with type 2 diabetes in European populations. The THADA gene is the target gene of a specific chromosomal rearrangement found in benign thyroid tumors, so it is also the most commonly used target gene in chromosomal rearrangement in benign epithelial tumors. In addition, the THADA gene may be associated...

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Problems solved by technology

In addition, THADA gene may be associated with polycystic ovary syndrome, prostate cancer, breast cancer

[0003] At present, THADA gene mutation detection methods mainly include: matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) technology, fluorescence quantitative PCR technology, Illumina fiber optic bead chip technology, matrix-assisted laser desorption ionization time-of-flight mass spectrometry technology is a Soft ionization technology has powerful and mature functions in the detection of protein and other biological macromolecules, but in the field of nucleic acid detection, due to the particularity of nucleic acid molecules, the detection is subject to certain restrictions

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

Although the Illumina fiber optic bead chip technology is a high-throughput detection system with high sensitivity and accuracy, it has a low degree of automation and many manual operations, which cannot meet the needs of practical applications.

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