On-Site Detection
of Enterocytozoon hepatopenaei in Shrimp: A Rapid and Reliable Colorimetric PSR
Method
Tran Hong Diem, Tran Thi Hau, Vo Thi Tu Binh, Le Thi Kim Dung and Phung Thi Thu
Huong
Res. J. Biotech.; Vol. 20(9); 109-117;
doi: https://doi.org/10.25303/209rjbt1090117; (2025)
Abstract
Enterocytozoon hepatopenaei (EHP) is a microsporidian parasite that significantly
threatens global shrimp farming, particularly affecting species like Penaeus monodon
and Penaeus vannamei. Early detection of EHP is crucial for effective disease control
in aquaculture environments. This study aimed to develop a rapid, sensitive and
specific diagnostic method for identifying EHP in shrimp populations through an
isothermal and direct colorimetric polymerase spiral reaction (PSR) assay. The PSR
assay was optimized for performance, yielding the best results at 65°C for 40 minutes.
We validated the specificity of our PSR primer set by testing it against a range
of aquatic pathogens, confirming no crossreactivity.
Furthermore, the limit of detection with a 95% probability was remarkably low, at
just 7.142 copies of the target gene per reaction. Clinical trials on shrimp samples
yielded encouraging results, with the direct colorimetric PSR method achieving 100%
sensitivity and specificity compared to traditional qPCR assays. Notably, the PSR
assay eliminates the need for DNA extraction, facilitating on-site detection at
shrimp farms. In summary, the newly developed colorimetric PSR assay offers a quick
and reliable option for detecting EHP, enhancing disease management in shrimp aquaculture
through its simplicity, accuracy and cost-effectiveness.
