Related Experiment Video
Updated: Mar 18, 2026
09:36
Open-Source Miniature Fluorimeter to Monitor Real-Time Isothermal Nucleic Acid Amplification Reactions in Resource-Limited Settings
Published on: February 3, 2021
5.4K
Comparison of fluorescent intercalating dyes for quantitative loop-mediated isothermal amplification (qLAMP)
Igor P Oscorbin1,2, Ekaterina A Belousova1, Aleksandr I Zakabunin1
1Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, Russia.
Biotechniques
|July 13, 2016
Summary
Quantitative loop-mediated isothermal amplification (qLAMP) relies on fluorescent dyes for pathogen detection. SYTO-82 dye offers the best performance, combining optimal time-to-threshold and signal-to-noise ratio for accurate results.
Area of Science:
- Molecular biology
- Biotechnology
- Analytical chemistry
Background:
- Quantitative loop-mediated isothermal amplification (qLAMP) is crucial for pathogen detection in biological and environmental samples.
- Accurate detection relies on sensitive fluorescent intercalating dyes.
- Optimizing dye selection is key to improving qLAMP assay performance.
Purpose of the Study:
- To comparatively evaluate the performance of six fluorescent intercalating dyes in qLAMP assays.
- To identify the most effective dye for real-time qLAMP applications.
- To provide data for selecting optimal dyes for pathogen detection.
Main Methods:
- Six fluorescent intercalating dyes (SYTO-9, SYTO-13, SYTO-82, SYBR Green I, SYBR Gold, EvaGreen) were tested.
- Dye performance was assessed across three distinct qLAMP model systems.
- Selected dyes underwent further enzyme and template titration studies.
Main Results:
- SYTO-9 and SYTO-82 exhibited superior performance compared to other tested dyes.
- SYTO-82 demonstrated the best overall performance, characterized by an optimal time-to-threshold (Tt) and signal-to-noise ratio (SNR).
- The findings highlight SYTO-82 as a highly effective dye for qLAMP.
Conclusions:
- SYTO-82 is the most effective fluorescent dye for quantitative loop-mediated isothermal amplification.
- This dye provides a robust combination of speed and signal clarity for pathogen detection.
- The study offers valuable insights for optimizing qLAMP protocols and enhancing diagnostic accuracy.
