Detection of Hematodinium in marine crustacean by loop-mediated isothermal amplification

Dinoflagellate in the genus of Hematodinium is an important parasite of marine crustacean.A rapid,convenient and reliable Hematodinium detection system was developed by using loop-mediated isothermal amplification(LAMP)method.A set of four primers including two outer primers and two inner primers were designed from the first internal transcribed space(rITS1)of Hematodiniumsp..LAMP assay amplifies Hematodiniumsp.'s DNA extracted from infected crabs under constant temperature of 65℃within 1h.The amplification products can be detected visually by observing whether white deposition comes into being or using SYBR green I dye,which turns green in the presence of amplified products and remains orange in its absence,or by electrophoresis.The LAMP assay was found to be 104 fold more sensitive than PCR assay.A comparison between LAMP and PCR assay using 25 clinical samples showed good correlation.The developed LAMP assay is simple,rapid,cost-effective,specific and highly sensitive for Hematodinium disease detection.