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

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

Publication date: Jan 20, 2024

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.

Concepts Keywords
Colorimetric Calcein
Expensive Colorimetric
Pcr COVID-19
Refugee Diagnosis
Rgb RT-LAMP
SARS-CoV-2

Semantics

Type Source Name
disease IDO assay
disease VO time
drug DRUGBANK Hydroxystilbamidine Isethionate
disease MESH COVID-19

Original Article

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