Probe library for detecting fusion gene of neurotrophin receptor kinase (NTRK) gene family, reagent, kit and application

Abstract

The invention provides a probe library for detecting a fusion gene of a neurotrophin receptor kinase (NTRK) gene family, a detection reagent comprising the probe library, application of the detectionreagent in preparing a kit used for detecting the fusion gene of the NTRK gene family, a corresponding kit and a detection method; the probe library comprises a first probe group and a second probe group, wherein detection of the fusion gene of the NTRK gene family is performed based on combination of a DNA interface and an RNA interface; the first probe group is used for DNA interface detection and capture; the second probe group is used for RNA interface detection and capture; the first probe group includes any one or more probes shown by SEQ ID NO.1-SEQ ID NO.170 of a nucleotide series; andthe second probe group includes any one or more probes shown by SEQ ID NO.21-SEQ ID.170 of the nucleotide series.

Description

Technical field [0001] The invention belongs to the field of biology, and specifically relates to a probe library for detecting NTRK gene family fusion genes, a detection reagent including the probe library, and application of the detection reagent in preparing a kit for detecting NTRK gene family fusion genes, and corresponding Kits and detection methods. Background technique [0002] NTRK is a gene encoding neurotrophic tyrosine receptor kinases. There are three genes in its family: NTRK1, NTRK2 and NTRK3 (NTRK1 / 2 / 3), which play an important role in maintaining the survival and normal function of the nervous system. The fusion of NTRK and other genes has been found to be related to the occurrence and development of a variety of tumors. Although the incidence of NTRK fusion in common tumors is less than 5%, in some rare tumors such as salivary gland tumors and infantile fibrous tumors, the NTRK fusion rate can reach more than 90%. In November 2018, the U.S. FDA approved Larotr...

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Problems solved by technology

It has the advantages of high stability, high sensitivity, good specificity, and short experimental cycle, but it can only detect one target at a time, requiring specialized tissue and multi-target detection. NTRK has 3 genes and needs to detect multiple positions Fusion requires the detection of multiple FISH probes. There are currently no commercial probes, which have significant false positive (FP) or false negative (FN), and can only be detected from the DNA level, and the detection cost is high; IHC Compared with the current molecular detection method, it is more economical and faster, but in IHC, it can only be detected at the protein level, and there is always a certain degree of subjectivity in the interpretation of the results. The judgment of weak positive results requires further use of FISH and other technologies. Verification; Fluorescent PCR technology is simple and easy to operate, short detection time, high sensitivity, good specificity, clear and intuitive judgment of results, wide application range, but only a few fusion genes of specific NTRK fusion partners can be detected, and only from Detection at the RNA level

Moreover, FISH and IHC cannot identify the gene (partner) involved in the fusion because they cannot know the detailed gene sequence, let alone know the specific fusion breakpoint and sequence information; while fluorescent PCR can only design probes and primers based on the known fusion Detected, but could not detect unknown fusions

[0005] High-throughput sequencing technology can sequence millions of short sequences in parallel, and has been widely used in the detection of tumor mutations. However, the current NGS method only judges DNA sequencing, which is likely to cause missed detection at the RNA level.

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