Detection of shrimp hemocyte iridescent virus by recombinase polymerase amplification assay.

摘要: Shrimp hemocyte iridescent virus (SHIV), which was first identified in white leg shrimp (Litopenaeus vannamei) China 2014, can cause extensive mortality and major economic losses the farming industry China. In this study, a novel real-time isothermal recombinase polymerase amplification (RPA) assay developed using TwistAmp exo kit for SHIV detection. First, five primers probe were designed capsid protein gene (GenBank: KY681039.1) according to TwistDx manual; next, optimal selected by comparison experiment. The specific did not react with spot syndrome (WSSV), infectious hypodermal hematopoietic necrosis (IHHNV), enterocytozoon hepatopenaei (EHP), macrobrachium rosenbergii nodavirus (MrNV) samples, as well pathogens of acute hepatopancreatic disease (AHPND). RPA reached detection limit 11 copies per reaction probit regression analysis. addition, detected positive plasmid samples at concentration 1000 copies/μL within 16.04 ± 0.72 min at single low operation temperature (39 °C). results proved that proposed method an accurate, sensitive, affordable, rapid tool be suitably applied diagnosis field conditions resource-poor settings.