Field deployable RPA-CRISPR detection tool

Antimicrobial resistance (AMR) represents a major global challenge, with water resources playing a critical role in the emergence and transmission of antibiotic-resistant bacteria and genes. Current laboratory-based methods for monitoring AMR in water resources are limited by high costs, lengthy turnaround times, and logistical challenges, which hinder timely interventions and effective mitigation strategies. This project aims to create a rapid, robust, and field-deployable diagnostic tool that integrates recombinase polymerase amplification (RPA) with CRISPR-based lateral flow assays (LFAs) for the detection of key AMR marker genes in various water sources. The project's approach leverages isothermal RPA for rapid (less than 15 minutes) amplification of target DNA, combined with highly specific and sensitive CRISPR-based detection methods. By incorporating these technologies into a user-friendly lateral flow format, the assay is designed to deliver results within 40-60 minutes, without the need for complex laboratory equipment.

Key methodologies include developing RPA-CRISPR assays that require minimal instrumentation and efficient sample preparation, as well as optimising assay performance for high sensitivity and specificity with real-world samples. We will validate these methods against standard molecular techniques such as qPCR. The anticipated outcome is a field-deployable diagnostic tool that enables the rapid detection of AMR genes, effectively addressing critical gaps in existing water monitoring practices. This tool is expected to significantly reduce logistical barriers, costs related to sample transport and processing, and diagnostic turnaround times, making AMR monitoring more accessible and scalable, especially for smaller or remote water utilities. Ultimately, this project aims to support timely interventions, implement effective AMR mitigation strategies, and enhance risk management of AMR threats across various sectors, including water, agriculture, aquaculture, and other sectors.

WEHI

Twelve months