Abstract
In this paper, we report a molecular diagnostic system-combining a colorimetric probe (RHthio-CuSO 4 ) for pyrophosphate sensing and isothermal gene amplification (ramified rolling circle amplification)-that operates with high selectivity and sensitivity for clinical point-of-care diagnosis of SARS-CoV-2. During the polymerase phase of the DNA amplification process, pyrophosphate was released from the nucleotide triphosphate as a side product, which was then sensed by our RHthio-CuSO 4 probe with a visible color change. This simple colorimetric diagnostic system allowed highly sensitive (1.13 copies/reaction) detection of clinical SARS-CoV-2 within 1 h, while also displaying high selectivity, as evidenced by its discrimination of two respiratory viral genomes (human rhino virus and respiratory syncytial virus) from that of SARS-CoV-2. All of the reactions in this system were performed at a single temperature, with positive identification being made by the naked eye, without requiring any instrumentation. The high sensitivity and selectivity, short detection time (1 h), simple treatment (one-pot reaction), isothermal amplification, and colorimetric detection together satisfy the requirements for clinical point-of-care detection of SARS-CoV-2. Therefore, we believe that this combination of a colorimetric probe and isothermal amplification will be useful for point-of-care testing to prevent the propagation of COVID-19.
Keywords: Colorimetric sensing; Isothermal amplification; Pyrophosphate; RHthio-CuSO 4 probe; Ramified rolling circle amplification (RAM); SARS-CoV-2.
【저자키워드】 SARS-CoV-2., isothermal amplification, Colorimetric sensing, Pyrophosphate, RHthio-CuSO 4 probe, Ramified rolling circle amplification (RAM), 【초록키워드】 COVID-19, Treatment, SARS-CoV-2, Genome, diagnostic, virus, point-of-care, amplification, colorimetric detection, sensitivity, respiratory syncytial virus, temperature, isothermal amplification, selectivity, molecular, Combination, nucleotide, Rolling circle amplification, Propagation, Pyrophosphate, isothermal, gene amplification, Reactions, naked eye, color change, reaction, this system, positive, DNA amplification, polymerase, probe, Prevent, respiratory viral, simple treatment, performed, displaying, released, evidenced, diagnosis of SARS-CoV-2, 【제목키워드】 COVID-19, Diagnosis,