Fabrication of graphene oxide-based pretreatment filter and Electrochemical-CRISPR biosensor for the field-ready cyanobacteria monitoring system
- Authors
- Kim, Gahyeon; Li, Yun Guang; Seo, Yoseph; Baek, Changyoon; Choi, Jin-Ha; Park, Hyunjun; An, Jeongyun; Lee, Myoungro; Noh, Seungwoo; Min, Junhong; Lee, Taek
- Issue Date
- Oct-2023
- Publisher
- Elsevier Ltd
- Keywords
- Aquatic ecosystem monitoring; Biosensor; Cyanobacteria; Hand-held filter; PCR-Free detection
- Citation
- Biosensors and Bioelectronics, v.237
- Journal Title
- Biosensors and Bioelectronics
- Volume
- 237
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/67768
- DOI
- 10.1016/j.bios.2023.115474
- ISSN
- 0956-5663
1873-4235
- Abstract
- Microcystis aeruginosa (M. aeruginosa) cause the eutrophication of lakes and rivers. To effectively control the overgrowth of M. aeruginosa, a suitable measurement method should be required in the aquatic fields. To address this, we developed a field-ready cyanobacterial pretreatment device and an electrochemical clustered regularly interspaced short palindromic repeats (EC-CRISPR) biosensor. The cyanobacterial pretreatment device consists of a syringe, glass bead, and graphene oxide (GO) bead. Then, the M. aeruginosa dissolved in the freshwater sample was added to fabricated filter. After filtration, the purified gene was loaded onto a CRISPR-based electrochemical biosensor chip to detect M. aeruginosa gene fragments. The biosensor was composed of CRISPR/Cpf1 protein conjugated with MXene on an Au microgap electrode (AuMGE) integrated into a printed circuit board (PCB). This AuMGE/PCB system maximizes the signal-to-noise ratio, which controls the working and counter electrode areas requiring only 3 μL samples to obtain high reliability. Using the extracted M. aeruginosa gene with a pre-treatment filter, the CRISPR biosensor showed a limit of detection of 0.089 pg/μl in fresh water. Moreover, selectivity test and matrix condition test carried out using the EC-CRISPR biosensor. These handheld pre-treatment kit and biosensors can enable field-ready detection of CyanoHABs. © 2023 Elsevier B.V.
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