Detection of SARS-CoV-2 Using Reverse Transcription Helicase Dependent Amplification and Reverse Transcription Loop-Mediated Amplification Combined with Lateral Flow Assay
Aleksandra Anna Zasada,
Ewa Mosiej,
Marta Prygiel,
Maciej Polak,
Karol Wdowiak,
Kamila Formińska,
Robert Ziółkowski,
Kamil Żukowski,
Kasper Marchlewicz,
Adam Nowiński,
Julia Nowińska,
Waldemar Rastawicki,
Elżbieta Malinowska
Affiliations
Aleksandra Anna Zasada
Department of Sera and Vaccines Evaluation, National Institute of Public Health NIH—National Research Institute, Chocimska 24, 00-791 Warsaw, Poland
Ewa Mosiej
Department of Sera and Vaccines Evaluation, National Institute of Public Health NIH—National Research Institute, Chocimska 24, 00-791 Warsaw, Poland
Marta Prygiel
Department of Sera and Vaccines Evaluation, National Institute of Public Health NIH—National Research Institute, Chocimska 24, 00-791 Warsaw, Poland
Maciej Polak
Department of Sera and Vaccines Evaluation, National Institute of Public Health NIH—National Research Institute, Chocimska 24, 00-791 Warsaw, Poland
Karol Wdowiak
Department of Sera and Vaccines Evaluation, National Institute of Public Health NIH—National Research Institute, Chocimska 24, 00-791 Warsaw, Poland
Kamila Formińska
Department of Sera and Vaccines Evaluation, National Institute of Public Health NIH—National Research Institute, Chocimska 24, 00-791 Warsaw, Poland
Robert Ziółkowski
The Chair of Medical Biotechnology, Faculty of Chemistry, Warsaw University of Technology, 00-664 Warsaw, Poland
Kamil Żukowski
Centre for Advanced Materials and Technologies CEZAMAT, Warsaw University of Technology, Poleczki 19, 02-822 Warsaw, Poland
Kasper Marchlewicz
The Chair of Medical Biotechnology, Faculty of Chemistry, Warsaw University of Technology, 00-664 Warsaw, Poland
Adam Nowiński
2nd Dept of Respiratory Medicine, National Institute of Tuberculosis and Lung, 01-138 Warsaw, Poland
Julia Nowińska
Faculty of Medicine, Medical University of Warsaw, 02-091 Warsaw, Poland
Waldemar Rastawicki
Department of Bacteriology and Biocontamination Control, National Institute of Public Health NIH—National Research Institute, 00-791 Warsaw, Poland
Elżbieta Malinowska
The Chair of Medical Biotechnology, Faculty of Chemistry, Warsaw University of Technology, 00-664 Warsaw, Poland
Rapid and accurate detection and identification of pathogens in clinical samples is essential for all infection diseases. However, in the case of epidemics, it plays a key role not only in the implementation of effective therapy but also in limiting the spread of the epidemic. In this study, we present the application of two nucleic acid isothermal amplification methods—reverse transcription helicase dependent amplification (RT-HDA) and reverse transcription loop-mediated amplification (RT-LAMP)—combined with lateral flow assay as the tools for the rapid detection of SARS-CoV-2, the etiological agent of COVID-19, which caused the ongoing global pandemic. In order to optimize the RT-had, the LOD was 3 genome copies per reaction for amplification conducted for 10–20 min, whereas for RT-LAMP, the LOD was 30–300 genome copies per reaction for a reaction conducted for 40 min. No false-positive results were detected for RT-HDA conducted for 10 to 90 min, but false-positive results occurred when RT-LAMP was conducted for longer than 40 min. We concluded that RT-HDA combined with LFA is more sensitive than RT-LAMP, and it is a good alternative for the development of point-of-care tests for SARS-CoV-2 detection as this method is simple, inexpensive, practical, and does not require qualified personnel to perform the test and interpret its results.
