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Title: A sensitive and specific fluorescent RT-LAMP assay for SARS-CoV‑2 detection in clinical samples
Authors: Ooi, Kean Hean
Liu, Mengying Mandy
Moo, Jia Rong
Nimsamer, Pattaraporn
Payungporn, Sunchai
Kaewsapsak, Pornchai
Tan, Meng How
Keywords: Engineering::Bioengineering
Issue Date: 2022
Source: Ooi, K. H., Liu, M. M., Moo, J. R., Nimsamer, P., Payungporn, S., Kaewsapsak, P. & Tan, M. H. (2022). A sensitive and specific fluorescent RT-LAMP assay for SARS-CoV‑2 detection in clinical samples. ACS Synthetic Biology, 11(1), 448-463.
Project: 2017-T1-001-214
Journal: ACS Synthetic Biology
Abstract: The raging COVID-19 pandemic has created an unprecedented demand for frequent and widespread testing to limit viral transmission. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) has emerged as a promising diagnostic platform for rapid detection of SARS-CoV-2, in part because it can be performed with simple instrumentation. However, isothermal amplification methods frequently yield spurious amplicons even in the absence of a template. Consequently, RT-LAMP assays can produce false positive results when they are based on generic intercalating dyes or pH-sensitive indicators. Here, we report the development of a sensitive RT-LAMP assay that leverages on a novel sequence-specific probe to guard against spurious amplicons. We show that our optimized fluorescent assay, termed LANTERN, takes only 30 min to complete and can be applied directly on swab or saliva samples. Furthermore, utilizing clinical RNA samples from 52 patients with COVID-19 infection and 21 healthy individuals, we demonstrate that our diagnostic test exhibits a specificity and positive predictive value of 95% with a sensitivity of 8 copies per reaction. Hence, our new probe-based RT-LAMP assay can serve as an inexpensive method for point-of-need diagnosis of COVID-19 and other infectious diseases.
ISSN: 2161-5063
DOI: 10.1021/acssynbio.1c00538
Rights: © 2022 American Chemical Society. All rights reserved.
Fulltext Permission: none
Fulltext Availability: No Fulltext
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