| 초록 |
We describe the integration of a novel isothermal amplification as nicking and extension chain reaction system-based amplification (NESBA) and digital real-time PCR (Dr. PCR) system for COVID-19 detection. The NESBA, an ultrasensitive version of NASBA, is nicking enzyme-based isothermal amplification by applying the primers labeled with a nicking site. The NESBA was integrated into the Dr. PCR system, implementing all the steps to realize digital analysis including the partition of nL-sized microarrays, isothermal amplification, and miniaturized real-time monitoring systems. By conducting parallel NESBA reactions in the localized 20,163 microarrays, SARS-CoV-2 gRNA was successfully identified down to 1 copy within a sample. Due to the intrinsic advantages including the ultrasensitive detection and absolute quantification, as well as the increase in the portability of the equipment, the digital real-time NESBA (Dr. NESBA) serves as a promising system to achieve POCT. |
