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Development of a recombinase polymerase amplification (RPA) fluorescence assay for the detection of enterocytozoon hepatopenaei (EHP)

文献类型： 外文期刊

作者： Li, Gen 1 ; Cong, Feng 2 ; Cai, Weiyou 1 ; Li, Jinhui 1 ; Wu, Miaoli 2 ; Xiao, Li 2 ; Hu, Xiaoliang 4 ; Zeng, Weiwei 5 ; He, 1 ;

作者机构： 1.South China Agr Univ, Coll Vet Med, Guangzhou 510640, Peoples R China

2.Guangdong Lab Anim Monitoring Inst, Guangzhou 510633, Peoples R China

3.Prov Key Lab Lab Anim, Guangzhou 510633, Guangdong, Peoples R China

4.Yibin Univ, Sch Life Sci & Food Engn, Yibin Key Lab Zool Divers & Ecol Conservat, Yibin 644000, Peoples R China

5.Foshan Univ, Sch Life Sci & Engn, Guangdong Prov Key Lab Anim Mol Design & Precise, Foshan 52823, Peoples R China

6.Chinese Acad Fishery Sci, Pearl River Fisheries Res Inst, Key Lab Fishery Drug Dev, Minist Agr,Key Lab Aquat Anim Immune Technol, Guangzhou 510380, Guangdong, Peoples R China

关键词： Recombinase polymerase amplification; Enterocytozoon hepatopenaei; Detection

期刊名称：AQUACULTURE REPORTS （ 影响因子：3.216； 五年影响因子：3.812 ）

ISSN： 2352-5134

年卷期： 2021 年 19 卷

页码：

收录情况： SCI

摘要： 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 degrees C. The detection limit of this EHP RPA was 10 copies/mu L of DNA molecules per reaction. The specificity of EHP RPA was tested, and the assay did not crossreact 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.

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