A Fast, Visual, and Instrument-free Platform Involving Rapid DNA Extraction, Chemical Heating, and Recombinase Aided Amplification for On-Site Nucleic Acid Detection

Xiao Fu Wang; Wen Qiang Chen; Jian Li Guo; Cheng Peng; Xiao Yun Chen; Xiao Li Xu; Wei Wei; Lei Yang; Jian Ca; Jun Feng Xu

doi:10.3389/fbioe.2021.764306

A Fast, Visual, and Instrument-free Platform Involving Rapid DNA Extraction, Chemical Heating, and Recombinase Aided Amplification for On-Site Nucleic Acid Detection

Front Bioeng Biotechnol. 2021 Nov 22:9:764306. doi: 10.3389/fbioe.2021.764306. eCollection 2021.

Authors

Xiao Fu Wang^{1

2}, Wen Qiang Chen^{1

3}, Jian Li Guo¹, Cheng Peng^{1

2}, Xiao Yun Chen^{1

2}, Xiao Li Xu^{1

2}, Wei Wei¹, Lei Yang¹, Jian Ca³, Jun Feng Xu^{1

2}

Affiliations

¹ State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Zhejiang Academy of Agricultural Sciences, Hangzhou, China.
² Key Laboratory of Information Traceability for Agricultural Products, Ministry of Agriculture of China, Hangzhou, China.
³ College of Biological and Food Engineering, Fuyang Normal University, Fuyang, China.

Abstract

The nucleic acid-based technique has been widely utilized in many fields including for on-site detection. However, traditional molecular detection techniques encounter limitations like relying on instruments, time consuming or complex operation, and cannot meet the demands of on-site testing. In this study, a rapid DNA extraction method (RDEM), recombinase aided amplification (RAA), and chemical heating packet (CHP) are integrated and termed as RRC platform for on-site detection of nucleic acid. For demonstration purposes, SHZD32-1 (a new transgenic soybean line from China) was detected using the novel platform to demonstrate its feasibility and capability for on-site detection. Using the RDEM, high-quality DNA appropriate for molecular detection was quickly extracted in 3-5 min. The heat energy generated by CHP was met the temperature requirements of RAA. Using the RRC platform, the whole detection process can be accomplished within only 30 min, and the results can be visually detected with glasses under blue light. No special or expensive instrument was needed for the detection process. This study provides a novel approach for on-site detection of nucleic acids besides providing valuable insight on related future research.

Keywords: RAA; chemical heating; nucleic acid; on-site detection; rapid DNA extraction.