Rapid and sensitive detection of iridovirus by loop-mediated isothermal amplification (LAMP)

Abstract

Red seabream iridovirus (RSIV), a member of the Iridoviridae family, is the causative pathogen of some of the most explosive epidemics of emerging viral diseases in many Asian countries, leading to huge economic losses in aquaculture. Rapid molecular detection for surveillance or diagnosis has been a critical component in reducing the prevalence of RSIV infection. In the present study, a novel and highly specific loop-mediated isothermal amplification (LAMP) assay for the sensitive and rapid detection of RSIV infection in fishes was developed. Using a set of synthesized primers matching a specific region of the RSIV genome (GenBank accession no.: AB666336.1), the efficiency and specificity of the LAMP assay were optimized in terms of the reaction temperature and DNA polymerase concentration, as they are the main determinants of the sensitivity and specificity of the LAMP assay. In particular, we demonstrated that our assay could be applied to efficiently detect RSIV infection in red sea bream. Our results provide a simple and convenient method for the detection of viral infection in aquatic organisms.