Fusion gene nucleic acid detection method based on combination of capillary electrophoresis fragment analysis and first-generation sequencing

Abstract

The invention discloses a fusion gene nucleic acid detection method based on combination of capillary electrophoresis fragment analysis and first-generation sequencing. The invention provides a kit for detecting multiple fusion genes. The kit comprises multiple pairs of amplification primer pairs, wherein each pair of amplification primers consists of a primer A and a primer B; the primer A sequentially consists of a linker sequence A used for combining a sequencing primer and a specific sequence A specifically combined with a target sequence of the fusion gene from the 5' end; the primer B sequentially consists of a linker sequence B for adjusting the length of an amplification product and a specific sequence B specifically combined with a target sequence of the fusion gene from the 5' end; and the 5' end of the primer A is modified with a fluorophore. By combining the capillary electrophoresis fragment analysis and the first-generation sequencing, fusion gene nucleic acid-based multiple detection and sequence synchronous acquisition are realized, so that while fragment analysis-based multiple detection is performed, first-generation sequencing is performed on a positive fusion gene to obtain a specific sequence of the positive fusion gene.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a fusion gene nucleic acid detection method based on the combination of capillary electrophoresis fragment analysis and first-generation sequencing. Background technique [0002] Fusion genes formed by chromosomal mutations usually play a key driving role in the occurrence and progression of tumors. More and more studies have shown that the detection and identification of fusion genes is of great importance for the diagnosis, treatment, prognosis evaluation and subsequent diagnosis of tumors. Lesion residual disease monitoring is of great significance. Current methods for detecting fusion genes usually include fluorescence in situ hybridization (FISH), real-time fluorescent PCR and next-generation high-throughput sequencing, etc., but these methods have their own defects or disadvantages. FISH technology has the characteristics of complicated operation, high labor cost a...

AI Technical Summary

Problems solved by technology

Using capillary fragment analysis alone can only obtain the fragment size, but not the specific sequence information; while the next generation sequencing method can obtain the sequence, but usually only for a single nucleic acid fragment target, and the operation steps are cumbersome

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