Facile, rapid, economical, and selective detection of antibiotic resistance genes from environment

Research Project, 2024 – 2026

Extensive worldwide use of antibiotics for humans and livestock has promoted spreading of antibiotic resistant bacteria and antibiotic resistance genes (ARGs) in the environment. Tighter control measures are needed for the release of ARGs in wastewater treatment plant effluents, water streams leaving the plants, and aquatic environments. The current methods used for the detection of ARGs are based on PCR, which makes them time consuming and costly as they require multi-step reactions, many reagents, trained personnel and complex instrumentation. In this project we will develop a graphene-based chip that uses ARG-targeting capacity of clustered regularly interspaced short palindromic repeats (CRISPR)-associated protein 9 (Cas9). Cas9 will be complexed with a specific ARG single-guide RNA and immobilized on graphene chip to yield on-chip electrical detection of ARGs. The output signal can be measured rapidly (in several minutes, much faster than PCR that takes several hours in a specialized lab) and simply (chip plugged via a USB port into a portable potentiometer). This method will enable fast, simple, economical, and specific detection of target ARG sequences within unamplified intact plasmid material collected from environmental samples. This fast detection method can help for the prompt action on application of efficient treatments for antibiotic resistance genes removal thereby improving the environment and protecting aquatic ecosystems.