A method for preparing double-stranded probes for detecting fusion genes and probes prepared by the method

Abstract

The invention relates to a preparation method of a fused gene capture probe and the probe prepared by adopting the method. The method comprises the steps that 1, a probe zone is determined according to an intron and an exon occurred at a target gene fusion site; 2, the probe zone is used as a template to design the probe zone, amplified from a genome DNA, of a primer; 3, a biological avidin label is added to the amplified probe zone; 4, gel extraction recovery is conducted on the probe zone added with the biological avidin label; 5, the probe zone added with the biological avidin label is randomly cut into 60-120 bp of fragment probes; 6, quantitative probe detection is performed. The method has the advantages of wide application range, short period and low cost. The probe prepared by adopting the method has the advantages that multiple coverages are achieved through random fragmentation, the probes in different zones are freely combined, efficient capture is achieved, and results are accurate.

Description

technical field [0001] The invention belongs to the fields of gene detection and molecular genetics, and in particular relates to a preparation method of a double-strand probe for detecting fusion genes. Background technique [0002] Fusion gene detection is of great significance to the diagnosis and treatment of many cancers. Currently, the main methods for fusion gene detection include fluorescence in situ hybridization and real-time quantitative polymerase chain reaction (QPCR). Fluorescence in situ hybridization is the gold standard for fusion gene detection. It has the advantage of being accurate, but for low-frequency fusions, it is easy to miss detection, the operation is complicated, and the requirements for operators are high. It can only determine the specific sequence of the approximate fusion site but not the site. QPCR detection detects fusion genes from the RNA level, which has high sensitivity, low cost, and short cycle. However, it can only detect fusion gene...

AI Technical Summary

Problems solved by technology

Fluorescence in situ hybridization is the gold standard for detection of fusion genes. Its advantage is that it is accurate, but it is easy to miss detection for low-frequency fusion, the operation is complicated, and the requirements for operators are high. It can only determine the specific sequence of the approximate fusion site but cannot locate the site.

QPCR detection detects fusion genes from the RNA level, which has high sensitivity, low cost, and short cycle, but it can only detect fusion gene types with known fusion sequences, and cannot detect unknown fusion types, nor can it determine the sequence of fusion sites

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