A Rapid COVID-19 RT-PCR Detection Assay for Low Resource Settings

Abstract

Quantitative reverse transcription polymerase chain reaction (RT-qPCR) assay is the gold standard recommended to test for acute SARS-CoV-2 infection. It has been used by the Centers for Disease Control and Prevention (CDC) and several other companies in their Emergency Use Authorization (EUA) assays. RT-qPCR requires expensive equipment such as RNA isolation instruments and real-time PCR thermal cyclers, which are not available in many low resource settings and developing countries. As a pandemic, COVID-19 has quickly spread to the rest of the world. Many underdeveloped and developing counties do not have the means for fast and accurate COVID-19 detection to control this outbreak. Using COVID-19 positive clinical specimens, we demonstrated that RT-PCR assays can be performed in as little as 12 minutes using untreated samples, heat-inactivated samples, or extracted RNA templates. Rapid RT-PCR was achieved using thin-walled PCR tubes and a setup including sous vide immersion heaters/circulators. Our data suggest that rapid RT-PCR can be implemented for sensitive and specific molecular diagnosis of COVID-19 in situations where sophisticated laboratory instruments are not available.

Tools and Techniques for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)/COVID-19 Detection

“…For developing countries or other resource-limited settings where instrumentation is lacking, other cost-sparing testing alternatives are being explored. Arumugam et al demonstrated a proof of principle of an RT-PCR that could be conducted in 12 min using a setup consisting of thin-walled PCR tubes, sous vide immersion heaters/circulators, and an endpoint readout performed with a light-emitting diode (LED) gel-viewing box ( 326 ). Such creative and innovative solutions from industry and academic settings help meet the global needs for SARS-CoV-2 laboratory testing, besides other NAATs rapidly being developed and validated.…”

End-to-end system for rapid and sensitive early-detection of SARS-CoV-2 for resource-poor and field-test environments using a $51 lab-in-a-backpack

“…In addition to the marginal cost per sample, for many lower-volume sites, one has to also consider the additional cost of infrastructure and staff training. Low-cost approaches have been advocated for both RT-LAMP and RT-qPCR tests [ 29 ].…”

“…These range from extremely low-cost ($10) device sets that could be operated even without electricity [ 30 ], to more expensive $300 equipment that can be used to run concurrent tests in 96-well plates and then reading out fluorescent results with a UV source and a smart phone or a tablet [ 29 ]. The appropriate (fixed capital) cost for such instruments depends strongly on the required number of tests to be run at a time, longevity and stable operation of the equipment, and situation-specific factors.…”