TERT gene mutation detection specific primers and liquid chip

Abstract

The invention discloses a TERT gene mutation detection liquid chip and specific primers. The liquid chip mainly comprises the following parts: each ASPE primer contains a tag sequence at the 5' terminal and a specific primer sequence aiming at target gene mutation locus at the 3' terminal, and the specific primer sequences are as below: a SEQ ID NO.15 and a SEQ ID NO.16 aiming at a T174G locus, a SEQ ID NO.17 and a SEQ ID NO.18 aiming at a C98T locus, a SEQ ID NO.19 and a SEQ ID NO.20 aiming at a G162A locus, a SEQ ID NO.21 and a SEQ ID NO.22 aiming at a G168A locus, a SEQ ID NO.23 and a SEQ ID NO.24 aiming at a C209T locus, a SEQ ID NO.25 and a SEQ ID NO.26 aiming at a G151T locus, and / or a SEQ ID NO.27 and a SEQ ID NO.28 aiming at an A129T locus; microballoons coated with anti-tag sequences; and amplification primers. The detection results from the detection liquid chip provided by the invention are 100% identical with those from a sequencing method, so as to realize parallel detection on wild types and mutants of a plurality of mutation loci.

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 TERT gene mutation detection and a liquid phase chip. Background technique [0002] TERT is called telomerase reverse transcriptase (Telomerase reverse transcriptase), which is located on chromosome 5 5P13.3. The TERT gene is a ribonucleoprotein located at the end of telomeres. Both are required for replication and for suppressing cellular senescence, and dysfunctional telomeres can affect gene stability and thereby increase the risk of tumor formation. The TERT gene encodes the catalytic subunit of telomerase, which is involved in the infinite proliferation of human cells and the pathogenesis of cancer cells. It is also a key gene in the development of cancer and is related to the occurrence of many cancers. It is related to basal cell carcinoma, lung cancer, bladder cancer, and prostate cancer. Cancer, cervical cance...

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Problems solved by technology

[0003] At present, the detection methods of TERT gene mutation mainly include: microarray technology, fluorescent quantitative PCR technology and matrix-assisted laser desorption ionization time-of-flight mass spectrometry technology (MALDI-TOF-MS) , microarray technology has problems such as high cost, complexity, low detection sensitivity, poor repeatability, narrow analysis range, complex synthesis and immobilization of probes, etc. In addition, because the hybridization is located on the surface of the solid phase, there is a certain degree of steric hindrance.

Fluorescent quantitative PCR technology has the disadvantages of low sensitivity, easy sample contamination, and high false positive rate. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry technology is a soft ionization technology that has powerful and mature functions in the detection of proteins and other biological macromolecules. However, in the field of nucleic acid detection, due to the particularity of nucleic acid molecules, the detection is subject to certain restrictions.

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