A loop mediated isothermal amplification (LAMP) method for detection of infectious hematopoietic necrosis virus (IHNV) in rainbow trout (Oncorhynchus mykiss)

A reverse transcription loop-mediated isothermal amplification (RT-LAMP) protocol was developed for detection of infectious hematopoietic necrosis virus (IHNV) RNA in rainbow trout (Oncorhynchus mykiss). A set of four primers, two outer and two inner primers for the RT-LAMP and the LAMP assay, were designed based on the sequence of G-protein of IHNV. Time and temperature conditions were optimized for 60 min at 63 degrees C for both RT-LAMP and LAMP protocols. The detection limit was found to be similar for both RT-LAMP and LAMP. When the sensitivity of RT-LAMP and LAMP were compared with conventional nested PCR, a 10-fold higher sensitivity was seen for the LAMP protocols.