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Probe group, kit and method for detecting NTRK fusion mutation

A technology of fusion mutation and probe set, applied in biochemical equipment and methods, DNA/RNA fragments, recombinant DNA technology, etc., can solve the problems of singleness, lack of comprehensiveness, difficulty in large-scale development, etc., and improve the quality of life , high detection sensitivity and wide coverage

Inactive Publication Date: 2020-03-06
苏州璞瑞卓越生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Furthermore, for the FISH method, a group of FISH separation probes is usually only used for the fusion mutation detection of one gene, which is relatively single and lacks comprehensiveness, making it difficult to carry out on a large scale.

Method used

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  • Probe group, kit and method for detecting NTRK fusion mutation
  • Probe group, kit and method for detecting NTRK fusion mutation
  • Probe group, kit and method for detecting NTRK fusion mutation

Examples

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Comparison scheme
Effect test

Embodiment

[0034] The source of samples in this example is the blood samples of 6 patients who complained of non-small cell lung cancer (all patients have signed the informed consent). The following is the genetic testing process of the 6 patients.

[0035] 1. Extraction of DNA

[0036] Whole blood samples using QIAamp TM DNA Mini Kit (Qiagen.Cat.no.51304) kit was used to extract nucleic acid samples from patients. The specific extraction process is:

[0037] 1. Pipette 20μL of QIAGEN Protease (Protease K100mg / mL) into the bottom of a sterile 2mL centrifuge tube;

[0038] 2. Add 200 μL whole blood sample to the above centrifuge tube, then add 4 μL RNase A (ribonuclease A 100 mg / mL), mix well by pipetting, incubate at room temperature for 5 minutes, then add 200 μL Buffer AL (lysis buffer), vortex Mix for 15 seconds;

[0039] 3. Place the centrifuge tube in a 56°C water bath and incubate for 10 minutes;

[0040] 4. Centrifuge briefly, add 200 μL ethanol (96-100%), vortex and mix for...

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Abstract

The invention discloses a probe group for detecting NTRK fusion mutation. The probe group comprises one or more probes of which the nucleotide sequences are shown as SeqID NO. 1-146. The invention also discloses a kit for detecting NTRK fusion mutation. The kit comprises the probe group. The invention further provides a method for detecting the NTRK fusion mutation by using the probe group. The method for detecting the NTRK fusion mutation by using the probe group designed by the invention shortens the hybridization time, and can detect whether the NTRK gene of a non-small cell lung cancer patient is subjected to fusion mutation or not more quickly. Compared with a detection gold standard FISH method, the method has the advantages of high detection sensitivity, wide coverage, high flux andthe like, and is beneficial to large-scale clinical development.

Description

technical field [0001] The invention belongs to the technical field of gene detection, and in particular relates to a probe set, a kit and a method for NTRK fusion mutation detection. Background technique [0002] NTRK (NeuroTrophin Receptor Kinase) is a neurotrophin receptor tyrosine kinase. The tropomyosin receptor kinase (TRK) family includes three proteins, TRKA, TRKB and TRKC, which are encoded by NTRK1, NTRK2 and NTRK3 genes, respectively, and these proteins are usually expressed in nerve tissue. Fusion mutations in the NTRK gene promote tumor formation. Dysregulation of the TRK pathway can lead to neurodevelopmental disorders and various central nervous system (CNS) diseases, but also induces cancer when the NTRK gene is fused to an unrelated gene. The World Health Organization estimates that NTRK fusions are present in 0.2% of NSCLC patients. The TRK fusion protein will be in a continuously active state, triggering a permanent signaling cascade that drives the spr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6827C12N15/11
CPCC12Q1/6827C12Q2531/113C12Q2535/122
Inventor 张惠丹戴敬刘琦赵洪玉
Owner 苏州璞瑞卓越生物科技有限公司
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