| Info. | Vol.17 - No.2 (2023.06.20) |
|---|---|
| Title | Paper-Based Radial Flow Assay Integrated to Portable Isothermal Amplification Chip Platform for Colorimetric Detection of Target DNA |
| Authors | Tai-Yong Kim 1,2, Sanha Kim 3, Jae Hwan Jung3, Min-Ah Woo 1 Tai-Yong Kim and Sanha Kim contributed equally to this work. *Min-Ah Woo mawoo@kfri.re.kr |
| Institutions | 1Research Group of Food Safety and Distribution, Korea
Food Research Institution, Wanju-Gun, Jeollabuk-do 55365, Republic of Korea
2Department of Food Science and Technology, Jeonbuk National University, Jeonju-si, Jeollabuk-do 54896,Republic of Korea 3Department of Pharmaceutical Engineering, Dankook University, Cheonan-si,Chungcheongnam-do 31116, Republic of Korea |
| Abstract | A novel integrated detection system that introduces a paper-chip-based molecular detection strategy into a polydimethylsi- loxane (PDMS) microchip and temperature control system was developed for on-site colorimetric detection of DNA. For the paper chip-based detection strategy, a padlock probe DNA (PLP)-mediated rolling circle amplification (RCA) reaction for signal amplification and a radial flow assay according to the Au-probe labeling strategy for visualization were optimized and applied for DNA detection. In the PDMS chip, the reactions for ligation of target-dependent PLP, RCA, and labeling were performed one-step under isothermal temperature in a single chamber, and one drop of the final reaction solution was loaded onto the paper chip to form a radial colorimetric signal. To create an optimal analysis environment, not only the optimization of molecular reactions for DNA detection but also the chamber shape of the PDMS chip and temperature control system were successfully verified. Our results indicate that a detection limit of 14.7 nM of DNA was achieved, and non-specific DNAs with a single-base mismatch at the target DNA were selectively discriminated. This integrated detection system can be applied not only for single nucleotide polymorphism identification, but also for pathogen gene detection. The adoption of inexpensive paper and PDMS chips allows the fabrication of cost-effective detection systems. Moreover, it is very suitable for operation in various resource-limited locations by adopting a highly portable and user-friendly detection method that minimizes the use of large and expensive equipment. |
| Keyword | DNA detection, Radial flow assay, Rolling circle amplification, PDMS chip,Paper chip, Portable platform |
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