Development of a recombinase polymerase amplification (RPA) fluorescence assay for the detection of enterocytozoon hepatopenaei (EHP)
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 sh...
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doaj-32ad72b7f35e4fe1981e85962e1167082021-03-01T04:15:41ZengElsevierAquaculture Reports2352-51342021-03-0119100584Development of a recombinase polymerase amplification (RPA) fluorescence assay for the detection of enterocytozoon hepatopenaei (EHP)Gen Li0Feng Cong1Weiyou Cai2Jinhui Li3Miaoli Wu4Li Xiao5Xiaoliang Hu6Weiwei Zeng7Dongsheng He8College of Veterinary Medicine of South China Agricultural University, Guangzhou, 510640, ChinaGuangdong laboratory animals monitoring institute and Guangdong, Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510633, ChinaCollege of Veterinary Medicine of South China Agricultural University, Guangzhou, 510640, ChinaCollege of Veterinary Medicine of South China Agricultural University, Guangzhou, 510640, ChinaGuangdong laboratory animals monitoring institute and Guangdong, Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510633, ChinaGuangdong laboratory animals monitoring institute and Guangdong, Provincial Key Laboratory of Laboratory Animals, Guangzhou, 510633, ChinaSchool of life sciences and food engineering, Yibin University, Yibin Key Laboratory of Zoological Diversity and Ecological Conservation, Yibin, 644000, China; Corresponding authors.School of Life Science and Engineering, Foshan University, Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding, Foshan University, Foshan, 52823, China; Key Laboratory of Fishery Drug Development, Ministry of Agriculture, Key Laboratory of Aquatic Animal Immune Technology, Guangdong Province, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, 510380, China; Corresponding author at: School of Life Science and Engineering, Foshan University, Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding, Foshan University, Foshan, 52823, China.College of Veterinary Medicine of South China Agricultural University, Guangzhou, 510640, China; Corresponding authors.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.http://www.sciencedirect.com/science/article/pii/S2352513420306773Recombinase polymerase amplificationEnterocytozoon hepatopenaeiDetection |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Gen Li Feng Cong Weiyou Cai Jinhui Li Miaoli Wu Li Xiao Xiaoliang Hu Weiwei Zeng Dongsheng He |
spellingShingle |
Gen Li Feng Cong Weiyou Cai Jinhui Li Miaoli Wu Li Xiao Xiaoliang Hu Weiwei Zeng Dongsheng He Development of a recombinase polymerase amplification (RPA) fluorescence assay for the detection of enterocytozoon hepatopenaei (EHP) Aquaculture Reports Recombinase polymerase amplification Enterocytozoon hepatopenaei Detection |
author_facet |
Gen Li Feng Cong Weiyou Cai Jinhui Li Miaoli Wu Li Xiao Xiaoliang Hu Weiwei Zeng Dongsheng He |
author_sort |
Gen Li |
title |
Development of a recombinase polymerase amplification (RPA) fluorescence assay for the detection of enterocytozoon hepatopenaei (EHP) |
title_short |
Development of a recombinase polymerase amplification (RPA) fluorescence assay for the detection of enterocytozoon hepatopenaei (EHP) |
title_full |
Development of a recombinase polymerase amplification (RPA) fluorescence assay for the detection of enterocytozoon hepatopenaei (EHP) |
title_fullStr |
Development of a recombinase polymerase amplification (RPA) fluorescence assay for the detection of enterocytozoon hepatopenaei (EHP) |
title_full_unstemmed |
Development of a recombinase polymerase amplification (RPA) fluorescence assay for the detection of enterocytozoon hepatopenaei (EHP) |
title_sort |
development of a recombinase polymerase amplification (rpa) fluorescence assay for the detection of enterocytozoon hepatopenaei (ehp) |
publisher |
Elsevier |
series |
Aquaculture Reports |
issn |
2352-5134 |
publishDate |
2021-03-01 |
description |
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. |
topic |
Recombinase polymerase amplification Enterocytozoon hepatopenaei Detection |
url |
http://www.sciencedirect.com/science/article/pii/S2352513420306773 |
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