Multiplex PCR primer composition, reagent and control system for next-generation sequencing of whole exons of TERT gene

Abstract

The invention provides a multiplex PCR primer composition, a reagent and a control system for next-generation sequencing of whole exons of TERT gene. The multiplex PCR primer composition for the second-generation sequencing of the whole exons of the TERT gene comprises primers shown as SEQ ID NO: 01 to SEQ ID NO: 50. The multiplex PCR primer composition, the reagent and the control system for thesecond-generation sequencing of the whole exons of the TERT gene provided by the invention solve the following existing technical problems: 1, the point mutation of a TERT gene promoter is mainly detected in the prior art, and the detection range is limited; 2, real-time fluorescent quantitative PCR detection is carried out, and the gene chip can only detect known sites; 3, the Sanger sequencing test length is in the range of less than 800bp; 4, when multiple sites are detected, the operation is tedious, and the cost is high; and 5, low-frequency mutation cannot be detected.

Description

technical field [0001] The invention relates to the field of gene sequencing, in particular to a multiplex PCR primer composition, reagent and control system for second-generation sequencing of whole exons of TERT gene. Background technique [0002] TERT (telomerase reverse transcriptase, telomerase reverse transcriptase) refers to the catalytic subunit with reverse transcriptase activity in telomerase, which is an important catalytic part of telomerase and affects the regulation mechanism of telomeres, thereby maintaining cell Immortalization and carcinogenesis. The TERT gene is located on chromosome 5, and its transcript NM_001193376.1 has 15 exons, with a total length of 3275 bp. TERT gene mutations are found in malignant tumors such as melanoma, lipoma, hepatoma, and glioma All have a certain detection rate, and the prognosis of positive patients is mostly poor, and they are biomarkers of related tumors. [0003] Regarding the mutation of the TERT gene, it is known tha...

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Problems solved by technology

[0004] Most of the TERT gene mutation detection in the prior art is to detect the C228T and C250T sites of the TERT gene promoter activity, which has the following limitations: 1. Real-time fluorescent quantitative PCR detection and gene chip detection cannot detect unknown sites; 2. The detection site of Sanger sequencing is limited to within 800bp; 3. When multiple sites are involved, the original technology is cumbersome and expensive; 4. It cannot detect low-frequency mutations

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