Advantages:
- Rapid results with minimal equipment: Delivers positive results in 10-20 minutes using only a simple heat block, with a distinct pink-to-yellow color change visible to the naked eye - significantly faster and simpler than standard RT-PCR
- Dual-target specificity and sensitivity; employs two distinct primer sets, achieving detection limits as low as 10 copies/µL
- Versatile surveillance applications across wastewater, agricultural, and envirnmental samples
Summary:
Effective detection of viral pathogens in environmental and agricultural contexts is critical for managing crop losses and monitoring public health risks. Traditional methods, such as ELISA or PCR often require expensive, specialized laboratory equipment and extensive sample preparation, making rapid, field-based detection challenging. Additionally, these techniques are associated with issues such as limited sensitivity, risk of nonspecific amplification or contamination, and lengthy turnaround times, particularly when viral concentrations are low or when complex matrices such as wastewater or plant tissues are involved.
Our inventors have developed two novel RT-LAMP (Reverse-Transcription Loop-Mediated Isothermal Amplification) assays for detecting PMMoV: -one targeting the viral RNA-dependent RNA polymerase gene and the other the coat protein gene- to achieve rapid, highly sensitive amplification at a constant 62°C in a simple 25 µL reaction. These assays produce a simple color change from pink to yellow upon amplification, enabling easy, field-based detection without requiring complex thermocyclers or fluorescence readers. With reliable detection of as few as 100 copies per µL for the RdRp target and 1000 copies per µL for the CP target and positive results in as little as 10-20 minutes, this approach significantly enhances speed compared to quantitative RT-PCR or ELISA methods. These assays serve as a robust, cost-effective, and versatile solution for monitoring PMMoV in agricultural and environmental samples, as well as for wastewater surveillance applications.
PMMoV LAMP assay produces a color change from pink to yellow upon amplification, with detection limits as low as 10 copies per µL for the RdRp target and 100 copies per µL for the CP target.
Desired Partnerships:
- License
- Sponsored Research
- Co-Development