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Updated: Aug 4, 2026
Multiplexed Isothermal Amplification Based Diagnostic Platform to Detect Zika, Chikungunya, and Dengue 1
Published on: March 13, 2018
PI primers increase the efficacy of LAMP and RT-LAMP for SARS-CoV-2 and MS2 phage detection
Igor P Oscorbin1, Lidiya M Novikova1, Evgeniy A Khrapov1
1Laboratory of Pharmacogenomics, Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, 8 Lavrentiev Avenue, 630090 Novosibirsk, Russia.
Abstract:
LAMP (Loop-mediated isothermal amplification) is a popular method for the molecular diagnostics of numerous pathogens, specifically useful for point-of-care testing. However, the efficacy and sensitivity of LAMP still need to be maximised for the best performance in clinical settings. Adding a novel fourth primer pair is a promising way to accelerate the LAMP speed. Here, we report PI primers that are part of inner primers and can be used in LAMP without a specific design. PI primers were tested in quantitative LAMP detecting SARS-CoV-2 and MS2. The new primers have increased the speed and sensitivity of quantitative LAMP, RT-LAMP, and duplex LAMP with artificial templates and RNA samples from nasal swabs. Adding PI primers could become a valuable option for LAMP optimisation, especially when a desirable LAMP target is a highly variable DNA sequence with a few conservative sites for primers.
Insights
Novel PI primers enhance the speed and sensitivity of Loop-mediated isothermal amplification (LAMP) assays. This optimization is particularly beneficial for molecular diagnostics, improving performance in clinical settings.
Area of Science:
- Molecular Biology
- Biotechnology
- Medical Diagnostics
Background:
- Loop-mediated isothermal amplification (LAMP) is a widely used molecular diagnostic technique, especially for point-of-care testing.
- Current LAMP methods require optimization to maximize efficacy and sensitivity for clinical applications.
- Accelerating LAMP speed and enhancing sensitivity are key goals for improving diagnostic performance.
Purpose of the Study:
- To introduce and evaluate a novel fourth primer pair, termed PI primers, for Loop-mediated isothermal amplification.
- To assess the impact of PI primers on the speed and sensitivity of quantitative LAMP assays.
- To demonstrate the utility of PI primers in various LAMP formats, including RT-LAMP and duplex LAMP.
Main Methods:
- PI primers were designed as a novel addition to existing LAMP primer sets, functioning as inner primers.
- Quantitative LAMP assays were performed using PI primers to detect SARS-CoV-2 and MS2 targets.
- The performance of PI primers was evaluated with both artificial templates and clinical RNA samples from nasal swabs.
- RT-LAMP and duplex LAMP assays were also conducted to assess the versatility of PI primers.
Main Results:
- The addition of PI primers significantly increased the speed of quantitative LAMP reactions.
- PI primers demonstrably enhanced the sensitivity of quantitative LAMP, RT-LAMP, and duplex LAMP assays.
- Improved performance was observed across artificial templates and clinical RNA samples, indicating broad applicability.
- The PI primers proved effective even with highly variable DNA targets possessing limited conserved regions.
Conclusions:
- PI primers represent a valuable tool for optimizing Loop-mediated isothermal amplification assays.
- This novel primer strategy offers a method to accelerate LAMP speed and improve sensitivity, especially for challenging targets.
- The integration of PI primers can enhance the diagnostic capabilities of LAMP, particularly in resource-limited or point-of-care settings.
