Construction and optimization of a LAMP-based diagnostic platform for acute hepatopancreatic necrosis disease in Penaeus vannamei.

Acute hepatopancreatic necrosis disease (AHPND) poses a major threat to global shrimp aquaculture, especially impacting Penaeus vannamei. Given the high cost of prevention and the absence of effective treatments, the most efficient way to control AHPND is through early detection to quickly identify the pathogen. This study presents the development and optimization of a low-cost, portable diagnostic platform utilizing loop-mediated isothermal amplification (LAMP) for the rapid detection of AHPND. This platform integrates fluorescence detection with a smartphone-compatible device, providing a convenient and effective solution for on-site diagnosis. We also screened fluorescent nucleic acid dyes with optimal adaptability and developed an efficient, user-friendly tool to improve the performance of the isothermal amplification method. In addition, a deep learning-based infection detection algorithm was developed for automated diagnosis. The visualization technology showed high specificity and sensitivity for detecting AHPND in shrimp samples, with a detection limit of 1 copies/µL. These results highlight the potential of this method in resource-limited environments and provide a valuable tool for early detection and management of AHPND in shrimp aquaculture.