Detection of SARS-CoV-2 Using Reverse Transcription Helicase Dependent Amplification and Reverse Transcription Loop-Mediated Amplification Combined with Lateral Flow Assay
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 pres...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-09-01
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| Series: | Biomedicines |
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| Online Access: | https://www.mdpi.com/2227-9059/10/9/2329 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Aleksandra Anna Zasada |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-10T00:38:29Z |
| format | Article |
| id | doaj.art-9601071bbc85410e9934846b44a3e9c3 |
| institution | Directory Open Access Journal |
| issn | 2227-9059 |
| language | English |
| last_indexed | 2024-03-10T00:38:29Z |
| publishDate | 2022-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomedicines |
| spelling | doaj.art-9601071bbc85410e9934846b44a3e9c32023-11-23T15:13:01ZengMDPI AGBiomedicines2227-90592022-09-01109232910.3390/biomedicines10092329Detection of SARS-CoV-2 Using Reverse Transcription Helicase Dependent Amplification and Reverse Transcription Loop-Mediated Amplification Combined with Lateral Flow AssayAleksandra Anna Zasada0Ewa Mosiej1Marta Prygiel2Maciej Polak3Karol Wdowiak4Kamila Formińska5Robert Ziółkowski6Kamil Żukowski7Kasper Marchlewicz8Adam Nowiński9Julia Nowińska10Waldemar Rastawicki11Elżbieta Malinowska12Department of Sera and Vaccines Evaluation, National Institute of Public Health NIH—National Research Institute, Chocimska 24, 00-791 Warsaw, PolandDepartment of Sera and Vaccines Evaluation, National Institute of Public Health NIH—National Research Institute, Chocimska 24, 00-791 Warsaw, PolandDepartment of Sera and Vaccines Evaluation, National Institute of Public Health NIH—National Research Institute, Chocimska 24, 00-791 Warsaw, PolandDepartment of Sera and Vaccines Evaluation, National Institute of Public Health NIH—National Research Institute, Chocimska 24, 00-791 Warsaw, PolandDepartment of Sera and Vaccines Evaluation, National Institute of Public Health NIH—National Research Institute, Chocimska 24, 00-791 Warsaw, PolandDepartment of Sera and Vaccines Evaluation, National Institute of Public Health NIH—National Research Institute, Chocimska 24, 00-791 Warsaw, PolandThe Chair of Medical Biotechnology, Faculty of Chemistry, Warsaw University of Technology, 00-664 Warsaw, PolandCentre for Advanced Materials and Technologies CEZAMAT, Warsaw University of Technology, Poleczki 19, 02-822 Warsaw, PolandThe Chair of Medical Biotechnology, Faculty of Chemistry, Warsaw University of Technology, 00-664 Warsaw, Poland2nd Dept of Respiratory Medicine, National Institute of Tuberculosis and Lung, 01-138 Warsaw, PolandFaculty of Medicine, Medical University of Warsaw, 02-091 Warsaw, PolandDepartment of Bacteriology and Biocontamination Control, National Institute of Public Health NIH—National Research Institute, 00-791 Warsaw, PolandThe Chair of Medical Biotechnology, Faculty of Chemistry, Warsaw University of Technology, 00-664 Warsaw, PolandRapid 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.https://www.mdpi.com/2227-9059/10/9/2329RT-HDART-LAMPdiagnosticspoint-of-careSARS-CoV-2isothermal amplification |
| spellingShingle | 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 Detection of SARS-CoV-2 Using Reverse Transcription Helicase Dependent Amplification and Reverse Transcription Loop-Mediated Amplification Combined with Lateral Flow Assay Biomedicines RT-HDA RT-LAMP diagnostics point-of-care SARS-CoV-2 isothermal amplification |
| title | Detection of SARS-CoV-2 Using Reverse Transcription Helicase Dependent Amplification and Reverse Transcription Loop-Mediated Amplification Combined with Lateral Flow Assay |
| title_full | Detection of SARS-CoV-2 Using Reverse Transcription Helicase Dependent Amplification and Reverse Transcription Loop-Mediated Amplification Combined with Lateral Flow Assay |
| title_fullStr | Detection of SARS-CoV-2 Using Reverse Transcription Helicase Dependent Amplification and Reverse Transcription Loop-Mediated Amplification Combined with Lateral Flow Assay |
| title_full_unstemmed | Detection of SARS-CoV-2 Using Reverse Transcription Helicase Dependent Amplification and Reverse Transcription Loop-Mediated Amplification Combined with Lateral Flow Assay |
| title_short | Detection of SARS-CoV-2 Using Reverse Transcription Helicase Dependent Amplification and Reverse Transcription Loop-Mediated Amplification Combined with Lateral Flow Assay |
| title_sort | detection of sars cov 2 using reverse transcription helicase dependent amplification and reverse transcription loop mediated amplification combined with lateral flow assay |
| topic | RT-HDA RT-LAMP diagnostics point-of-care SARS-CoV-2 isothermal amplification |
| url | https://www.mdpi.com/2227-9059/10/9/2329 |
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