Accessible LAMP-Enabled Rapid Test (ALERT) for Detecting SARS-CoV-2

Area of Science:

• Biotechnology
• Molecular Biology
• Infectious Disease Diagnostics

Background:

• Current COVID-19 diagnostic methods like PCR and antigen tests have limitations including cost, turnaround time, lab dependency, and false negative rates.
• The pandemic exposed critical needs for accessible, scalable, and accurate population-level infection detection.
• Existing molecular amplification techniques often require complex laboratory infrastructure and trained personnel.

Purpose of the Study:

• To develop an inexpensive, simple, and rapid do-it-yourself (DIY) workflow for detecting viral RNA.
• To achieve high sensitivity and specificity for viral detection comparable to or exceeding existing methods.
• To create a versatile platform adaptable for various targets and point-of-care applications.

Main Methods:

• Developed a 60-minute workflow utilizing reverse transcription loop-mediated isothermal amplification (RT-LAMP) for viral RNA detection.
• Incorporated features like wax-encapsulated reagents for shelf-stability, sequence-specific QUASR for improved specificity, and magnetic concentration for enhanced sensitivity.
• Integrated a quality control target and enabled multiplexing for co-detection of other respiratory viruses.

Main Results:

• Achieved high sensitivity (0.1 to 2 viral particles/μL) and specificity (>97% true negative rate) for viral RNA detection.
• Demonstrated a 60-minute turnaround time for results from nasal swabs or saliva.
• The ALERT system showed increased shelf-life and ambient temperature storage capabilities.

Conclusions:

• The Accessible LAMP-Enabled Rapid Test (ALERT) provides a robust, sensitive, and specific molecular diagnostic solution.
• ALERT addresses key limitations of current infectious disease testing, offering a viable option for at-home and point-of-care use.
• The flexible design of ALERT allows for easy adaptation to detect various pathogens and nucleic acid markers beyond SARS-CoV-2.