The emerging microsporidian parasite Enterocytozoon hepatopenaei (EHP) causes retardation of shrimp growth, leading to significant financial losses in shrimp aquaculture. Therefore, the development of an efficient and sensitive detection method will be conducive to the prevention and control of the shrimp parasite. In this study, we developed and evaluated a rapid real-time recombinase polymerase amplification (RPA) method that can detect EHP within 15 min at a constant temperature of 38.5 °C. The detection limit of this EHP RPA was 10 copies/μL of DNA molecules per reaction. The specificity of EHP RPA was tested, and the assay did not cross-react with white spot syndrome virus (WSSV), shrimp hemocyte iridescent virus (SHIV), infectious hypodermal and hematopoietic necrosis virus (IHHNV), or Vibrio parahaemolyticus. Field and clinical applicability of this assay was evaluated using 61 field samples. The coincidence rate of the detection results for the clinical samples between RPA and qPCR was 95.1 %. In summary, the real-time RPA analysis provides an efficient and sensitive detection method for EHP.

新兴的小孢子虫寄生虫肝细胞戊型肝炎（EHP）导致虾生长迟缓，导致虾养殖的重大经济损失。因此，开发一种高效灵敏的检测方法将有利于虾寄生虫的防治。在这项研究中，我们开发并评估了一种快速实时重组酶聚合酶扩增（RPA）方法，该方法可以在38.5°C的恒定温度下于15分钟内检测EHP。该EHP RPA的检测限为每个反应10个拷贝/μLDNA分子。测试了EHP RPA的特异性，并且该测定未与白斑综合症病毒（WSSV），虾血细胞虹彩病毒（SHIV），传染性皮下和造血坏死病毒（IHHNV）或副溶血性弧菌发生交叉反应。。使用61个现场样品评估了该测定的现场和临床适用性。RPA与qPCR之间临床样品检测结果的符合率为95.1％。总之，实时RPA分析为EHP提供了一种有效而灵敏的检测方法。