Sugar production contributes more than $2 billion annually to the Australian economy, mainly through export earnings (DAFF 2019; QEAS 2019). However, the associated costs of disease management and prevention are up to 40% of the economic production value (Magarey et al. 2003). Therefore, effective and informed disease management and biosecurity planning/surveillance for endemic and exotic pathogens are critical for the ongoing profitability of the Australian sugarcane industry. Management requires early, fast, and accurate diagnostic and quantification of causative organisms and their epidemiology. While existing diagnostic methods are robust and well-established, they are time-consuming, expensive and laboratory-based. Here, we report on the development of a novel handheld diagnostic device, SugarPATH (Figure 1a), suitable for detecting target pathogens within 20 min. SugarPATH is designed to overcome the shortcomings in existing sugarcane pathogen diagnostics by providing Australia with a rapid and accurate diagnostic tool for early detection and to provide proper biosecurity surveillance and disease management. Key outputs Using loop-mediated isothermal amplification (LAMP) and colorimetric quantitative readout results, SugarPATH has been tested to detect Leifsonia xyli subsp. xyli, the causal bacteria of ratoon stunting disease in xylem sap. As demonstrated in Figure 1b, an ex situ LAMP analysis at different time intervals indicates a positive diagnosis at approximately 12 min. SugarPATH has been integrated with a novel single-step method for in situ nucleic acid isolation and quantification from diverse sample types (Chakraborty et al. 2023).