| 基于TaqMan qPCR检测凡纳滨对虾样品中虾肝肠胞虫并样检测方法的评价 |
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| 引用本文： | 宋增磊,董宣,赵若恒,王秀华,武和英,于党辉,谢国驷,黄捷. 基于TaqMan qPCR检测凡纳滨对虾样品中虾肝肠胞虫并样检测方法的评价[J]. 渔业科学进展, 2019, 40(3): 122-132 |
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| 作者姓名： | 宋增磊 董宣 赵若恒 王秀华 武和英 于党辉 谢国驷 黄捷 |
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| 作者单位： | 上海海洋大学水产与生命学院 上海 201306;中国水产科学研究院黄海水产研究所青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室农业农村部海水养殖病害防治重点实验室青岛市海水养殖流行病学与生物安保重点实验室 青岛 266071;中国水产科学研究院黄海水产研究所青岛海洋科学与技术试点国家实验室海洋渔业科学与食物产出过程功能实验室农业农村部海水养殖病害防治重点实验室青岛市海水养殖流行病学与生物安保重点实验室 青岛 266071;大连海洋大学水产与生命学院 大连 116023 |
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| 基金项目： | 现代农业产业技术体系(CARS-48)、中国东盟海上合作基金项目(2016~2018)和中国水产科学研究院基本科研业务费专项(2017HY-ZD10)共同资助 |
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| 摘 要： | 对山东海阳和潍坊的2个养殖凡纳滨对虾群体取样，采用TaqMan qPCR逐尾检测肝胰腺中的虾肝肠胞虫(Enterocytozoon hepatopenaei, EHP)载量，再将提取的DNA样品按5并1(5∶1)、25并1(25∶1)、50并1(50∶1)、100并1(100∶1)和150并1(150∶1)进行并样，检测并样的EHP载量。设定不同临界循环数为假定灵敏度，定性判断各单尾检测阳性及并样组阳性，比较不同并样模式与检测阳性率、诊断灵敏度、诊断特异性等之间的关系以及定量的准确性。结果表明，检测灵敏度过低，会降低高并样率检测的准确性；阳性率在30%以上时，高并样率的检测结果与单样品检测相符性很好；高载量感染的阳性率不低于6.7%时，50∶1以内的并样能准确得出检测结果；低载量感染的阳性率不低于16%时，25∶1以内的并样能得出较好结果；高载量感染的1.3%阳性率和低载量感染的8%阳性率可能导致所有并样出现假阴性结果；各种并样模式均有很好的诊断特异性，50∶1并样的诊断灵敏度与OIE标准推荐的5∶1并样的接近；各并样检测的EHP载量与单样品检测平均值之比在0.27~2.83范围，二者存在极显著相关性，各种并样模式的定量检测结果在数量级水平能大致反映样品的平均EHP载量。本研究为水生动物疫病诊断和流行病学调查的样品检测提供了参考依据。
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| 关 键 词： | 凡纳滨对虾 虾肝肠胞虫(EHP) qPCR 并样检测 |
| 收稿时间： | 2018-04-21 |
| 修稿时间： | 2018-05-04 |
Evaluation on the detection of Enterocytozoon hepatopenaei in pooled DNA samples of Litopenaeus vannamei based on TaqMan qPCR |
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| SONG Zenglei,DONG Xuan,ZHAO Ruoheng,WANG Xiuhu,WU Heying,YU Danghui,XIE Guosi and HUANG Jie. Evaluation on the detection of Enterocytozoon hepatopenaei in pooled DNA samples of Litopenaeus vannamei based on TaqMan qPCR[J]. Progress in Fishery Sciences, 2019, 40(3): 122-132 |
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| Authors: | SONG Zenglei DONG Xuan ZHAO Ruoheng WANG Xiuhu WU Heying YU Danghui XIE Guosi HUANG Jie |
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| Affiliation: | College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306. 2. Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences; Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao); Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture and Rural Affairs; Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity; Qingdao 266071,Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences; Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao); Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture and Rural Affairs; Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity; Qingdao 266071,College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023,Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences; Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao); Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture and Rural Affairs; Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity; Qingdao 266071,College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306. 2. Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences; Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao); Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture and Rural Affairs; Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity; Qingdao 266071,College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306. 2. Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences; Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao); Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture and Rural Affairs; Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity; Qingdao 266071,Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences; Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao); Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture and Rural Affairs; Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity; Qingdao 266071 and College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306. 2. Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences; Laboratory for Marine Fisheries Science and Food Production Processes, Pilot National Laboratory for Marine Science and Technology (Qingdao); Key Laboratory of Maricultural Organism Disease Control, Ministry of Agriculture and Rural Affairs; Qingdao Key Laboratory of Mariculture Epidemiology and Biosecurity; Qingdao 266071 |
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| Abstract: | |
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| Keywords: | Litopenaeus vannamei Enterocytozoon hepatopenaei (EHP) qPCR Pooled sample detection |
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