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A universal fluorescence biosensor based on rolling circle amplification and locking probe for DNA detection.

Ying Fang¹, Lanxin Nie¹, Suqin Wang¹

¹College of Chemistry and Chemical Engineering, Jiangxi Normal University, Nanchang, 330022, P.R. China.

Mikrochimica Acta

|July 1, 2024

Summary

A novel biosensor utilizes rolling circle amplification (RCA) and DNA replacement for stable DNA signal detection. This universal DNA detection method shows promise for identifying genetic information in targets like human immunodeficiency virus (HIV).

Keywords:

Fluorescent detection HIV Homologous DNA of let-7a Locking probe

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Area of Science:

Biotechnology
Molecular Biology
Biosensor Technology

Background:

Developing sensitive and stable biosensors for DNA detection is crucial for diagnostics.
Existing methods may lack the sensitivity or universality required for diverse genetic targets.

Purpose of the Study:

To develop a stable DNA signal amplification sensor for universal DNA detection.
To demonstrate the biosensor's efficacy using human immunodeficiency virus (HIV) DNA and let-7a miRNA.

Main Methods:

Utilized target DNA-controlled rolling circle amplification (RCA).
Incorporated locking probe DNA replacement technology for signal generation.
Employed T4 DNA ligase for padlock probe cyclization and Phi29 DNA polymerase for RCA.

Main Results:

Achieved stable DNA signal amplification through RCA.
Demonstrated successful detection of target DNA fragments.
Showcased a fluorescence recovery mechanism regulated by DNA hybridization and strand displacement.

Conclusions:

The developed biosensor offers a universal platform for DNA detection.
The combination of RCA and DNA replacement technology provides a stable and sensitive detection system.
This approach has potential applications in genetic information analysis and disease diagnostics.