LAUSR

Abstract Enterocytozoon hepatopenaei (EHP) has caused mortality in Pacific white shrimp (Penaeus vannamei) and resulted in large economic losses in shrimp farming. The best way to avoid this parasitic disease is to find out about them early. In this study, we describe a novel, sensitive and specific rapid isothermal recombinase polymerase amplification assay (RPA) for detecting EHP. We evaluated the ability, including the specificity and sensitivity, of the EHP-RPA assay to detect EHP. By optimizing the reaction time and temperature, and screening primers based on the conserved small subunit ribosomal RNA gene, we determined the optimal reaction conditions. The RPA could be performed at 30 °C for 40 min without cross-reactivity with other pathogens. Sensitivity assay indicated that the limit of detection for EHP-RPA, PCR and real-time PCR was 8 × 102, 8 × 102 and 8 × 101 copies/μL, respectively. Using 18 samples collected from shrimp farms, we demonstrated that RPA enables simple, convenient and rapid EHP detection, especially when laboratory equipment is lacking. This new RPA method is easy to implement and has great potential for use in both laboratory and field testing applications.