Efficient detection of pathogen virus in sand dabs, Paralichthysolivaceususingloop-mediatedisothermalamplification (LAMP)

Abstract

Viral hemorrhagic septicaemia virus (VHSV) and Marine birnavirus (MABV) is the causative pathogen for 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 pathogens infection. A loop-mediated isothermal amplification (LAMP) of DNA is currently one of the most commonly used molecular diagnostic tools due to its simple, fast and easy to amplify target DNA under isothermal conditions. In the present study, a novel and highly specific LAMP assay for the sensitive and rapid detection of VHSV and MABV infection in fishes was developed. Using a set of synthesized primers matching a specific region of genome, 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 VHSV and MABV infection in Paralichthys olivaceus and other virus family. Our results provide a simple and convenient method for the detection of viral infection in aquatic organisms.apid molecular detection for surveillance or diagnosis has been a critical component in reducing the prevalence of pathogens infection. A loop-mediated isothermal amplification (LAMP) of DNA is currently one of the most commonly used molecular diagnostic tools due to its simple, fast and easy to amplify target DNA under isothermal conditions. In the present study, a novel and highly specific LAMP assay for the sensitive and rapid detection of VHSV and MABV infection in fishes was developed. Using a set of synthesized primers matching a specific region of genome, 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 VHSV and MABV infection in Paralichthys olivaceus and other virus family. Our results provide a simple and convenient method for the detection of viral infection in aquatic organisms.