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?:abstract
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The 2019 novel coronavirus (COVID-19) is a newly emerged strain that has never been found in humans before At present, the laboratory-based reverse transcription-polymerase chain reaction (RT-PCR) is the main method to confirm COVID-19 infection The intensification of the COVID-19 epidemic overwhelms limited clinical resources in particular, but not only, in developing countries, resulting in many patients not being tested for the infection and in large queues of potentially infected individuals waiting to be tested while providing a breeding ground for the disease We describe here a rapid, highly sensitive, point-of-care, molecular test amenable for use at home, in the clinic, and at points of entry by minimally trained individuals and with minimal instrumentation Our test is based on loomediated isothermal amplification (COVID-19 LAMP) and for higher sensitivity on nested nucleic acid, two stage isothermal amplification (COVID-19 Penn-RAMP) Both tests can be carried out in closed tubes with either fluorescence or colorimetric (e g , leuco crystal violet LCV) detection COVID-19 LAMperforms on par with COVID-19 RT-PCR COVID-19 RAMhas 10 fold better sensitivity than COVID-19 LAMand COVID-19 RT-PCR when testing purified targets and 100 times better sensitivity than COVID-19 LAMand COVID-19 RT-PCR when testing rapidly prepared sample mimics Due to fortunate scarcity of COVID-19 infections in the USA, we were not able to test our assays and methods with patient samples We hope that such tests will be carried out by colleagues in impacted countries Our Closed-Tube Penn-RAMhas the potential to significantly reduce false negatives while being amenable to use with minimal instrumentation and training
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