Developing a Point-of-Risk Molecular Test for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)

Developing a Point-of-Risk Molecular Test for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2)

Laura E. Lamb1,2, Sarah N. Bartolone1, Elijah Ward1, Michael B. Chancellor1,2

1 Department of Urology, Beaumont Health System, Royal Oak, Michigan, United States of America

2 Oakland University William Beaumont School of Medicine, Rochester Hills, Michigan, United States of America

Introduction/Objective:

Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), which causes coronavirus disease 2019 (COVID-19), has become a global health concern that requires a rapid and inexpensive diagnostic test. Quantitative reverse transcription PCR (qRT-PCR) is currently the gold standard for SARS-CoV-2 detection. However, Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) may allow for inexpensive and rapid testing outside the constraints of a traditional clinical laboratory. Our objective was to develop an RT-LAMP assay for detection of SARS-CoV-2.

Method/Description:

RT-LAMP primers were designed to a consensus sequence of SARS-CoV-2 strains available in January 2020. Several RT-LAMP primer sets were tested for performance. The final RT-LAMP primer set was tested in both simulated patient samples and clinical patient samples. Simulated samples were generated by spiking serum, urine, saliva, oropharyngeal swabs, and nasopharyngeal swabs with a portion of the SARS-CoV-2 nucleic sequence. RNA isolated from nasopharyngeal swabs collected from actual COVID-19 patients was also tested. The samples were tested using RT-LAMP as well as by conventional qRT-PCR. Specificity of the RT-LAMP was evaluated by also testing against other related coronaviruses.

Results:

RT-LAMP detected SARS-CoV-2 in both simulated patient samples and clinical specimens with similar sensitivity to PCR. RT-LAMP was performed in 30–45 minutes. RT-LAMP was specific to SAR-CoV-2 and did not detect genetically related viruses.

Take-home message/conclusions:

RT-LAMP is a novel method to detect SARS-CoV-2 rapidly and without expensive equipment or reagents. This approach could be used for monitoring of exposed individuals or potentially aid with screening efforts.

• Describe funding body that supported the work (if any)- This work was supported by the Maureen

and Ronald Hirsch family philanthropic contribution.

• Describe conflict of interest (if any) - None