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A new simplified sequence-dependent loop-mediated isothermal amplification (LAMP) detection method.
- Source :
-
Analytical & Bioanalytical Chemistry . Jul2024, Vol. 416 Issue 18, p4143-4152. 10p. - Publication Year :
- 2024
-
Abstract
- Fluorescence dye-based loop-mediated isothermal amplification (LAMP) is a sensitive nucleic acid detection method, but is limited to single-plex detection and may yield non-specific signals. In this study, we propose a bifunctional probe-based real-time LAMP amplification method for single-plexed or multiplexed detection. The bifunctional probe is derived by modifying the 5′ end of the fluorophore and an internal quencher on one of the LAMP primers; therefore, it can simultaneously be involved in the LAMP process and signal amplification. The fluorescence intensity undergoes a cumulative exponential increase during the incorporation of the bifunctional probe into double-stranded DNA amplicons. The bifunctional probe-based LAMP method is simplified and cost-effective, as the primer design and experimental operations align entirely with the ordinary LAMP. Different from other current probe-based methods, this method does not require additional enzymes, sequences, or special probe structures. Also, it is 10 min faster than several other probe-based LAMP methods. The bifunctional probe-based LAMP method allows the simultaneous detection of the target Vibrio parahaemolyticus DNA and the internal amplification control in a one-pot reaction, demonstrating its potential for multiplexed detection. [ABSTRACT FROM AUTHOR]
Details
- Language :
- English
- ISSN :
- 16182642
- Volume :
- 416
- Issue :
- 18
- Database :
- Academic Search Index
- Journal :
- Analytical & Bioanalytical Chemistry
- Publication Type :
- Academic Journal
- Accession number :
- 178445347
- Full Text :
- https://doi.org/10.1007/s00216-024-05340-7