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Enterospora epinepheli is a microsporidian parasite that poses a severe threat to the grouper farming industry, with limited options for prevention and treatment. Regular monitoring and early diagnosis are essential for managing infections. In this study, two rapid diagnostic methods were developed: a real-time enzymatic recombinase amplifi-cation (RT-ERA) assay and an enzymatic recombinase amplification combined with a lateral flow dipstick (ERA-LFD) assay, both targeting the 18S rDNA gene of E. epinepheli. These methods operate under isothermal conditions (≤40 ◦C) and offer rapid detection, with RT-ERA achieving results within 14~20 min and ERA-LFD within approximately 10 min. The detection limit for both methods was 2 × 100 copies/µL, with high specificity and no cross-reactivity with other aquaculture pathogens. Validation with grouper tissue and aquaculture water samples from Hainan, China, showed 100% concordance with basic ERA and superior performance compared to conventional PCR. These assays provide effective tools for early diagnosis, pathogen load quantification, and environmental monitoring, contributing to the prevention and control of E. epinepheli infections in aquaculture.

Doi: org/10.1016/j.aca.2023.341391