Issue 24, 2022

Specific detection of fusion transcripts based on a duplex-specific nuclease and isothermal exponential amplification reaction

Abstract

Fusion genes are mostly found in tumor tissues, but are in low expression levels in healthy tissues, making them good candidate biomarkers for tumor diagnosis and therapy. Here, we propose a duplex-specific nuclease-isothermal exponential amplification reaction (DSN-IEXPAR) method for the detection of fusion transcripts. A DNA probe is specifically designed for fusion transcript recognition and hybridization, and DSN cleavages the DNA probe in the DNA/RNA duplex. Through controlling the recognition and cleavage temperature, DSN can only cut the DNA probe fully matched with the target fusion transcript rather than other transcripts containing partial the same sequence, endowing the proposed method with high specificity to the fusion transcript in the presence of homologous sequences. The truncated DNA probe after cutting can subsequently trigger IEXPAR as a probe, so as low as 100 fM fusion transcript can be detected with the proposed DSN-IEXPAR. The evaluation of the analytical performance of DSN-IEXPAR demonstrates that it can provide an effective platform for fusion transcript detection in the ordinary laboratory and clinical diagnosis.

Graphical abstract: Specific detection of fusion transcripts based on a duplex-specific nuclease and isothermal exponential amplification reaction

Supplementary files

Article information

Article type
Paper
Submitted
15 ربيع الأول 1444
Accepted
07 ربيع الثاني 1444
First published
08 ربيع الثاني 1444

Analyst, 2022,147, 5649-5654

Specific detection of fusion transcripts based on a duplex-specific nuclease and isothermal exponential amplification reaction

F. Su, T. Li, X. He and Z. Li, Analyst, 2022, 147, 5649 DOI: 10.1039/D2AN01665F

To request permission to reproduce material from this article, please go to the Copyright Clearance Center request page.

If you are an author contributing to an RSC publication, you do not need to request permission provided correct acknowledgement is given.

If you are the author of this article, you do not need to request permission to reproduce figures and diagrams provided correct acknowledgement is given. If you want to reproduce the whole article in a third-party publication (excluding your thesis/dissertation for which permission is not required) please go to the Copyright Clearance Center request page.

Read more about how to correctly acknowledge RSC content.

Social activity

Spotlight

Advertisements