Publications
Functional & integrative genomicsJan 2024 |
24
(
1
),
16
DOI:
10.1007/s10142-024-01297-z

Ultrasensitive and fast detection of SARS-CoV-2 using RT-LAMP without pH-dependent dye

Alsaeed, Moneerah; Alhamid, Galyah; Tombuloglu, Huseyin; Kabanja, Juma H; Karagoz, Aysel; Tombuloglu, Guzin; Rabaan, Ali A; Al-Suhaimi, Ebtesam; Unver, Turgay
Product Used
Genes
Abstract
This study investigates the performance of reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the colorimetric detection of SARS-CoV-2 using fluorometric dye, namely, calcein. The detection limit (LoD) with the N-ID1 primer set resulted in superior performance, corresponding to ~ 2 copies/reaction or ~ 0.1 copies/μL of the RNA sample. The color development can be observed by the naked eye, using an ultraviolet (UV) transilluminator or a hand-UV light without the requirement of expensive devices. The average time-to-reaction (TTR) value was 26.2 min in high-copy number samples, while it was about 50 min in rRT-PCR. A mobile application was proposed to quantify the positive and negative results based on the three-color spaces (RGB, Lab, and HSB). Compared to rRT-PCR (n = 67), this assay allows fast and sensitive visual detection of SARS-CoV-2, with high sensitivity (90.9%), selectivity (100%), and accuracy (94.03%). Besides, the assay was sensitive regardless of variants. Since this assay uses a fluorescent dye for visual observation, it can be easily adapted in RT-LAMP assays with high sensitivity. Thus, it can be utilized in low-source centers and field testing such as conferences, sports meetings, refugee camps, companies, and schools.
Product Used
Genes

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