Five-color FISH probe system and method for detecting multiple genes at one time

Abstract

The invention discloses a five-color FISH probe system and method for detecting multiple genes at a time. The probe system is ingenious in design, the states of ALK, ROS1, RET and MET genes of the non-small cell lung cancer can be simultaneously detected on a specimen slice through single-time FISH, and therefore whether the fusion gene exists in a late-stage NSCLC patient or not is rapidly determined, specimens and reagents are saved, and the detection time is greatly shortened. The four genes ALK, ROS1, RET and MET are jointly detected, and the sensitivity and accuracy of prognosis prediction are improved. The corresponding molecular targeting drug is selected according to the detection result, and formulation of a clinical treatment scheme of lung cancer can be more reliably guided, so that prognosis is improved.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a five-color FISH probe system and method for one-time detection of multiple genes. Background technique [0002] Lung cancer ranks first in the mortality rate of malignant tumors in my country. Lung cancer can be divided into two categories: small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC). Non-small cell lung cancer accounts for about 85% of all lung cancer cases. The 5-year survival rate of lung cancer patients in my country is only 13%. If NSCLC, especially lung adenocarcinoma, cannot be detected with high sensitivity for gene mutations, the opportunity for precise targeted therapy will be lost, which will seriously affect the survival and quality of life of patients. [0003] At present, chemotherapy and radiotherapy are still one of the main means of NSCLC treatment. In the past few decades, although scientists have continuously explored NSCLC chemotherapy...

AI Technical Summary

Problems solved by technology

[0013] Although FISH is highly specialized, the use of FISH technology to detect multiple NSCLC driver genes still faces the following problems: First, because FISH is a pathological analysis at the cellular level, and most NSCLC are usually diagnosed at an advanced stage, its Most of the genetic testing specimens are lung biopsy specimens. After several rounds of slices such as conventional HE, immunohistochemistry, EGFR mutation detection, etc., the remaining biopsy tissues are often less and cannot meet the needs of individual screening; At present, most conventional FISH can only detect the fusion or amplification of one gene at a time, and the detection throughput is low. For multiple hot genes, FISH technology can only be used to continuously detect each driver gene one by one, which will lead to low work efficiency. It takes a long time to wait for the test results; finally, due to the relatively high price of FISH testing, the current method of testing multiple fusion genes in batches puts a certain amount of economic pressure on patients

[0014] For example, at present, the ROS1 gene, ALK gene, RET gene and MET gene are 4 probes respectively, and the color of each probe of these four genes on the market is double color (red / green or orange / green) ), at least 4 paraffin-embedded sections are required as samples, 4 experiments are performed, 4 reagents are consumed, and they are interpreted under a fluorescence microscope

Considering the high cost of testing and / or the small sample size available, patients lose the opportunity to detect abnormalities in four genes, thereby missing out on corresponding treatments

At present, there is no clinical kit that can detect the four hotspot genes of ALK, ROS1, RET and MET at the same time and at one time.

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