Primer, probe and detection reagent kit for detecting RET fusion gene

Abstract

The invention discloses a primer and a probe for detecting an RET fusion gene. The primer comprises an RET fusion mutation specificity reverse transcription primer, an RET gene fusion mutation detection specificity primer and a probe nucleotide sequence. The invention discloses a detection reagent kit for detecting the RET fusion gene. The detection reagent kit comprises an mRNA reverse transcription cDNA system, wherein the mRNA reverse transcription cDNA system is used for detecting the PCR reaction of the detection reagent. The specific primer and probe technology is adopted, so that the human RET gene fusion mutation can be specifically detected; a real-time fluorescent PCR system is established, and 17 fusion mutations of the RET genes can be simultaneously detected; the sensitivity is high, the mutation of 5 to 10 copies can be detected; the operation is simple, the detection is cheap, and the clinical application range is wide; the specimen detection range is wide, and the specimen can be fresh pathological tissues, paraffin-embedded tissues or pleural fluid; the detection speed is high, and the detection process can be completed in 90 minutes.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a primer, a probe and a detection kit for RET fusion gene detection. Background technique [0002] Lung cancer is the most common malignant tumor worldwide, and its morbidity and mortality rank first among all cancers. Every year, more than 1.3 million patients die of lung cancer worldwide, nearly half of which occur in developing countries. According to the statistics of the Ministry of Health of my country in 2010, the mortality rate of lung cancer was 30.83 / 100,000, and lung cancer has become the malignant tumor with the highest morbidity and mortality. (Orras JM, Fernandez E, Gonzalez JR, et al. Lung cancer mortality in European regions (1955-1997). Ann Oncol, 2003, 14(1): 159-161; Ministry of Health of the People's Republic of China, "2010 China Health Statistical Yearbook). [0003] Lung cancer can be divided into two categories based on histopathology: small cell lung can...

AI Technical Summary

Problems solved by technology

[0006] At present, the detection method of RET fusion mutation is mainly the fluorescence in situ hybridization (FISH) method, but the detection specificity of the FISH method is poor, the sensitivity is low, and the detection time is long.

In addition, FISH detection requires expensive special instruments and equipment, the cost of reagents is high, and the operation is complicated, which limits the widespread use of clinical detection.

FISH testing requires that the test samples should not be stored for too long. Using samples with a long storage time for testing will lead to false negatives and affect the accuracy of the test.

Therefore, the FISH detection method cannot meet the needs of clinicians for the practical application of detection. It is urgent to develop a highly sensitive detection method that can simultaneously detect multiple RET gene fusions, so as to quickly detect multiple RET fusion mutations at the same time. To provide scientific reference for clinical individualized treatment of lung cancer

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