Development of a rapid assay to detect the dinoflagellate Amyloodinium ocellatum using loop-mediated isothermal amplification (LAMP)

摘要: Amyloodinium ocellatum is a highly pathogenic dinoflagellate parasite with global distribution that causes high mortalities in the culture of tropical and sub-tropical marine estuarine fishes. Diagnosis typically occurs through gross examination following onset morbidity, at which point treatment limited benefit. In present study, new molecular diagnostic tool for rapid detection A. (AO) was developed using loop-mediated isothermal amplification method (LAMP). The AO-LAMP assay designed specific set four primers - two outer inner targeting six different regions on 5' end Small Subunit rDNA region (SSU rDNA) ocellatum. assay, optimized 25-30 min 62°C, amplified DNA from extracted both water gill tissue samples did not amplify closely related sp ecies. limit 10 fg, exceptionally higher than conventional PCR (1 pg). addition, standardized capable detecting single tomonts trophonts; affected by presence possible inhibitory substances environmental or samples. study provides novel useful simple, sensitive samples, could assist early improved control infections aquaculture systems.