Primer composition and reagent for fusion gene detection

Abstract

The present invention discloses a primer composition and a reagent for fusion gene detection. A human lung cancer fusion gene can be detected based on a multiplex PCR method. The primer composition can specifically amplify at least 50 fusion mutation forms of five fusion driving genes such as ALK, NRG, NTRK1, RET and ROS1 in an RNA sample through sequences as shown in SEQ ID NO:1-54, and can be used as a house-keeping gene for sample quality control. A reaction can be carried out in one tube, and can also be carried out in multiple tubes. During the multi-tube reaction, a specific probe composition modified with a fluorophore and having sequences as shown in SEQ ID NO 55-70 can be added to form the qPCR reagent. When the qPCR reagent is used for detecting a positive sample, the fusion genein 0.04 ng RNA can be reported. The process is simple and efficient, and the reagent is easy to operate and low in cost, and has a good prospect.

Description

technical field [0001] The invention relates to the technical field of gene detection, in particular to a primer composition and a reagent for fusion gene detection. Background technique [0002] Breakage and resplicing at the genome level often occur during tumorigenesis. When two genes are broken in half and spliced ​​together incorrectly, a new gene may be formed, which is called a fusion gene. When the carcinogenic fusion gene is transcribed into mRNA, its structure can be divided into two obvious parts: the 3' end usually comes from genes that have kinase activity and originally stimulate cell growth and proliferation, such as ALK, RET, etc. These genes are called fusion gene mutations The source of the sequence at the 5' end is more diverse, and its source gene is also called the "partner gene" of the fusion gene mutation. The common feature of these sequences is that they have a high expression in tumor cells and are not easily regulated. This leads to excessive act...

AI Technical Summary

Problems solved by technology

FISH is the current gold standard for fusion detection, but its sensitivity is low (10-15%), the operation is time-consuming and complicated, and it relies on human judgment

The advantage of IHC is that the price is lower, but the judgment criteria are more subjective, and it is impossible to directly detect fusion gene fragments

qPCR has high sensitivity (1-5%), strong operability, and low price. The disadvantage is that generally only one or similar known fusion mutations can be detected at a time

The sensitivity of NGS is slightly lower than that of qPCR. If the design is reasonable, there are more types of fusion mutations that can be detected at one time, but there are still problems of long time-consuming and high detection costs.

All in all, the existing detection methods have certain limitations and need to be improved.

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