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Rapid detection of Enterocytozoon hepatopenaei in shrimp through an isothermal recombinase polymerase amplification assay
Zhou, Shuhong1,3; Wang, Mengqiang1; Liu, Mei1; Jiang, Keyong1; Wang, Baojie1; Wang, Lei1,2,4
2020-05-15
发表期刊AQUACULTURE
ISSN0044-8486
卷号521页码:6
通讯作者Wang, Baojie(wangbaojie@qdio.ac.cn) ; Wang, Lei(leiwang@qdio.ac.cn)
摘要Enterocytozoon hepatopenaei (EHP) has caused mortality in Pacific white shrimp (Penaeus vannamei) and resulted in large economic losses in shrimp farming. The best way to avoid this parasitic disease is to find out about them early. In this study, we describe a novel, sensitive and specific rapid isothermal recombinase polymerase amplification assay (RPA) for detecting EHP. We evaluated the ability, including the specificity and sensitivity, of the EHP-RPA assay to detect EHP. By optimizing the reaction time and temperature, and screening primers based on the conserved small subunit ribosomal RNA gene, we determined the optimal reaction conditions. The RPA could be performed at 30 degrees C for 40 min without cross-reactivity with other pathogens. Sensitivity assay indicated that the limit of detection for EHP-RPA, PCR and real-time PCR was 8 x 10(2), 8 x 10(2) and 8 x 10(1) copies/mu L, respectively. Using 18 samples collected from shrimp farms, we demonstrated that RPA enables simple, convenient and rapid EHP detection, especially when laboratory equipment is lacking. This new RPA method is easy to implement and has great potential for use in both laboratory and field testing applications.
关键词Penaeus vannamei Enterocytozoon hepatopenaei Recombinase polymerase amplification
DOI10.1016/j.aquaculture.2020.734987
收录类别SCI
语种英语
资助项目National Key R&D Program of China[2019YFD0900401] ; National Natural Science Foundation of China-Joint Fund of Shandong People's Government[2017T3011]
WOS研究方向Fisheries ; Marine & Freshwater Biology
WOS类目Fisheries ; Marine & Freshwater Biology
WOS记录号WOS:000521282400061
出版者ELSEVIER
引用统计
被引频次:17[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.qdio.ac.cn/handle/337002/166997
专题实验海洋生物学重点实验室
通讯作者Wang, Baojie; Wang, Lei
作者单位1.Chinese Acad Sci, Inst Oceanol, CAS Key Lab Expt Marine Biol, Qingdao 266071, Peoples R China
2.Natl Lab Marine Sci & Technol, Lab Marine Biol & Biotechnol, Qingdao 266237, Peoples R China
3.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
4.Chinese Acad Sci, CAS Ctr Ocean Megasci, Qingdao 266400, Peoples R China
第一作者单位中国科学院海洋研究所
通讯作者单位中国科学院海洋研究所;  中国科学院海洋大科学研究中心
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GB/T 7714
Zhou, Shuhong,Wang, Mengqiang,Liu, Mei,et al. Rapid detection of Enterocytozoon hepatopenaei in shrimp through an isothermal recombinase polymerase amplification assay[J]. AQUACULTURE,2020,521:6.
APA Zhou, Shuhong,Wang, Mengqiang,Liu, Mei,Jiang, Keyong,Wang, Baojie,&Wang, Lei.(2020).Rapid detection of Enterocytozoon hepatopenaei in shrimp through an isothermal recombinase polymerase amplification assay.AQUACULTURE,521,6.
MLA Zhou, Shuhong,et al."Rapid detection of Enterocytozoon hepatopenaei in shrimp through an isothermal recombinase polymerase amplification assay".AQUACULTURE 521(2020):6.
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