Abstract
To safely re-open economies and prevent future outbreaks, rapid, frequent, point-of-need, SARS-CoV-2 diagnostic testing is necessary. However, existing field-deployable COVID-19 testing methods require the use of uncomfortable swabs and trained providers in PPE, while saliva-based methods must be transported to high complexity laboratories for testing. Here, we report the development and clinical validation of High-Performance Loop-mediated isothermal Amplification (HP-LAMP), a rapid, saliva-based, SARS-CoV-2 test with a limit of detection of 1.4 copies of virus per µl of saliva and a sensitivity and specificity with clinical samples of > 96%, on par with traditional RT-PCR based methods using swabs, but can deliver results using only a single fluid transfer step and simple heat block. Testing of 120 patient samples in 40 pools comprised of 5 patient samples each with either all negative or a single positive patient sample was 100% accurate. Thus, HP-LAMP may enable rapid and accurate results in the field using saliva, without need of a high-complexity laboratory.
Highlights
To safely re-open economies and prevent future outbreaks, rapid, frequent, point-of-need, SARSCoV-2 diagnostic testing is necessary
Reverse Transcription Loop-mediated isothermal Amplification (RT-LAMP), is a targeted isothermal nucleic acid amplification method that utilizes a combination of 2–3 primer sets and a DNA polymerase with high strand displacement activity[11]
While RT-LAMP has been used for SARS-CoV-2 detection by several groups[6,12,13,14], these methods require a prior extraction step or lengthy sample treatment, multiple fluid transfer steps, or lack the accuracy and limit of detection necessary for clinical implementation, and are not suitable for clinical testing outside of a laboratory
Summary
To safely re-open economies and prevent future outbreaks, rapid, frequent, point-of-need, SARSCoV-2 diagnostic testing is necessary. To develop HP-LAMP, we first designed novel primers for targeting the SARS-CoV-2 virus. Existing primers used for RT-PCR and LAMP based nucleic acid testing of SARS-CoV-2 target the GC-rich regions located at the 5′ and 3′ ends of the virus.
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