Direct on-the-spot detection of SARS-CoV-2 in patients

Author:

Ben-Assa Nadav1,Naddaf Rawi1,Gefen Tal1,Capucha Tal12,Hajjo Haitham134,Mandelbaum Noa1,Elbaum Lilach1,Rogov Peter5ORCID,King Daniel A6,Kaplan Shai7,Rotem Assaf8,Chowers Michal910,Szwarcwort-Cohen Moran11,Paul Mical12,Geva-Zatorsky Naama113ORCID

Affiliation:

1. Department of Cell Biology and Cancer Science, Technion Integrated Cancer Center (TICC),Rappaport Faculty of Medicine, Technion-Institute of Technology, Haifa 3525433, Israel

2. Department of Oral and Maxillofacial Surgery, Rambam Medical Care Center, Haifa 31999, Israel

3. Department of Immunology, Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa 3525433, Israel

4. Department of Neuroscience, Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa 3525433, Israel

5. Independent Researcher, Winchester, MA 01890, USA

6. Pulmonary and Respiratory Critical Care Division, Meir Medical Center, Kfar Saba 44281, Israel

7. Robiotec Ltd, Rehovot 7634709, Israel

8. Independent Reearcher, Newton, MA 02459, USA

9. Infectious Diseases Unit, Meir Medical Center, Kfar Saba 44281, Israel

10. Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv Isarel, 3200

11. Virology Laboratory, Rambam Health Care Campus, Haifa 31999, Israel

12. Division of Infectious Diseases, Rambam Health Care Center, and Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa 31999, Israel

13. MaRS Centre, Canadian Institute for Advanced Research (CIFAR) Azrieli Global Scholar, Toronto, ON M5G 1M1, Canada

Abstract

Many countries are currently in a state of lockdown due to the SARS-CoV-2 pandemic. One key requirement to safely transition out of lockdown is the continuous testing of the population to identify infected subjects. Currently, detection is performed at points of care using quantitative reverse-transcription PCR, thus requiring dedicated professionals and equipment. Here, we developed a protocol based on reverse transcribed loop-mediated isothermal amplification for the detection of SARS-CoV-2. This protocol is applied directly to SARS-CoV-2 nose and throat swabs, with no RNA purification step required. We tested this protocol on over 180 suspected patients, and compared the results to those obtained using the standard method. We further succeeded in applying the protocol to self-collected saliva samples from confirmed cases. Since the proposed protocol can detect SARS-CoV-2 from saliva and provides on-the-spot results, it allows simple and continuous surveillance of the community. Impact statement Humanity is currently experiencing a global pandemic with devastating implications on human health and the economy. Most countries are gradually exiting their lockdown state. We are currently lacking rapid and simple viral detections, especially methods that can be performed in the household. Here, we applied RT-LAMP directly on human clinical swabs and self-collected saliva samples. We adjusted the method to allow simple and rapid viral detection, with no RNA purification steps. By testing our method on over 180 human samples, we determined its sensitivity, and by applying it to other viruses, we determined its specificity. We believe this method has a promising potential to be applied world-wide as a simple and cheap surveillance test for SARS-CoV-2.

Publisher

SAGE Publications

Subject

General Biochemistry, Genetics and Molecular Biology

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