Primers, probes and kits for detecting fusion mutations of human alk gene and their detection methods

Abstract

The invention discloses a primer, a probe, a kit and a detection method for detecting fusion mutations of human ALK gene, and the primer and probe include: designing on the broken exon sequences of six partner genes in the ALK fusion gene Multiple upstream primers, shared downstream primers and fluorescent detection probes designed on ALK exon 20; 6 partner gene break exon sequences in ALK fusion gene include: EML4‑exon2 / 6 / 13 / 14 / 17 / 20, TFG ‑exon5, KIF5B‑exon15 / 17 / 24, TPR‑exon15, HIPI‑exon21, SEC31A‑exon21; the kit can cover up to 28 ALK fusion forms, with high specificity and sensitivity, and can effectively avoid false positives. The established one-step reverse transcription detection method can simplify operation and avoid contamination.

Description

technical field [0001] The invention relates to the field of in vitro molecular diagnosis, in particular to a primer, a probe, a kit and a detection method for detecting human ALK gene fusion mutation. Background technique [0002] In my country, the incidence of tumors is increasing rapidly. Lung cancer is a major killer that threatens human health, with high mortality and poor prognosis. The driving genes of lung cancer include EGFR mutation, ALK fusion gene, ROS1 gene rearrangement and c-MET gene amplification, etc. Among them, ALK fusion gene refers to the fusion of ALK gene (anaplastic lymphoma kinase, anaplastic lymphoma kinase) and echinoderm microtubule associated protein-like 4 (echinoderm microtubμle associated protein-like4, EML4) and other genes, the incidence rate in lung cancer patients About 3% to 7%. At present, multiple ALK gene fusion partner genes have been found, such as KIF5B, TFG, KLC1, PTPN3, HIP1 and TPR genes, among which EML4-ALK fusion accounts f...

AI Technical Summary

Problems solved by technology

However, most of the existing kits currently use the two-step RT-PCR method, which is prone to cross-contamination, and most of them only detect EML4-ALK fusion, while ignoring other important fusion sites, which is easy to cause missed detection, and false positives and false negatives The rate is high, the sensitivity is low, so it is difficult to promote in clinical practice; the market needs a test kit with many ALK fusion sites, high specificity and sensitivity, which can effectively avoid false positives, and the detection method can simplify operation and avoid pollution; solve such problems

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