Abstract
A probing system has been developed based on dual-site ligation-assisted loop-mediated isothermal amplification (dLig-LAMP) for the selective colorimetric detection of SARS-CoV-2. This approach can induce false-positive and -negative detection in real clinical samples; dLig-LAMP operates with improved selectivity. Unlike RT-LAMP, the selectivity of dLig-LAMP is determined in both the ligation and primer binding steps, not in the reverse transcription step. With this selective system in hand, we developed a colorimetric signaling system for point-of-care detection. We also developed a colorimetric probe for sensing pyrophosphate, which arises as a side product during the LAMP DNA amplification. Thus, dLig-LAMP appears to be an alternative method for improving the selectivity problems associated with reverse transcription. In addition, combining dLig-LAMP with colorimetric pyrophosphate probing allows point-of-care detection of SARS-CoV-2 within 1 h with high selectivity.
Keywords: Colorimetric detection; PP Probe; SARS-CoV-2; Splint R Ligase; dLig-LAMP.
【저자키워드】 SARS-CoV-2, colorimetric detection, PP Probe, Splint R Ligase, dLig-LAMP., 【초록키워드】 point-of-care, RT-LAMP, Loop-mediated isothermal amplification, LAMP, reverse transcription, isothermal amplification, selectivity, False-positive, binding, Signaling, Pyrophosphate, alternative method, problem, primer, hand, selective, DNA amplification, probe, approach, addition, appear, induce, arise, Splint, 【제목키워드】 point-of-care, isothermal amplification,