| 白斑综合征病毒环介导等温扩增快速检测方法的建立 |
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| 引用本文： | 何琳,徐海圣,王美珍,戎华南.白斑综合征病毒环介导等温扩增快速检测方法的建立[J].水产学报,2010,34(4):598-603. |
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| 作者姓名： | 何琳 徐海圣 王美珍 戎华南 |
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| 作者单位： | 1. 浙江大学动物科学学院,浙江,杭州,310029
2. 慈溪市水产技术推广中心,浙江,慈溪,315300 |
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| 基金项目： | 浙江省科技计划项目(2005C33053); 宁波市农业科技攻关项目(2008C10041); 宁波市海洋与渔业专项资金项目(7-3) |
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| 摘 要： | 根据对虾白斑综合征病毒(WSSV)囊膜蛋白VP28基因保守序列,利用Primer Explorerv 4.0软件设计了4条引物,建立了白斑综合征病毒环介导等温扩增快速检测方法,对反应温度和反应时间等参数进行了优化,同时将建立的LAMP检测方法与巢式PCR进行了比较分析。结果表明,LAMP最适反应在64℃恒温条件60min内完成,凝胶电泳呈现梯型条带;反应体系中添加SYBR Green I荧光染料后,绿色的阳性结果明显区别于橙色阴性结果。LAMP方法的最低检出限为100拷贝/μL,灵敏度较巢式PCR高100倍,而且LAMP方法在1h内即可完成检测,操作简单,无需复杂仪器,肉眼可直接观察检测结果。用建立的LAMP方法对临床发病南美白对虾样品进行了检测,结果表明,LAMP方法适合对虾白斑综合征病毒的现场快速检测。
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| 关 键 词： | 对虾 白斑综合征病毒 环介导等温扩增 快速检测 |
| 收稿时间： | 2009/12/15 0:00:00 |
| 修稿时间： | 2010/1/14 0:00:00 |
Development of rapid detection for white spot syndrome virus by loop-mediated isothermal amplification |
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| XU Haisheng,Wang Meizhen and Rong Huanan.Development of rapid detection for white spot syndrome virus by loop-mediated isothermal amplification[J].Journal of Fisheries of China,2010,34(4):598-603. |
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| Authors: | XU Haisheng Wang Meizhen and Rong Huanan |
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| Institution: | HE Lin1,XU Hai-sheng1,WANG Mei-zhen2,RONG Hua-nan2(1.College of Animal Science,Zhejiang University,Hangzhou 310029,China,2.Spreading Center of Technology in Aquatic Science of Cixi City,Zhejiang Province,Cixi 315300,China) |
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| Abstract: | A rapid,sensitive and simple method for white spot syndrome virus(WSSV) detection was established.Loop-mediated isothermal amplification(LAMP) assay is a novel method of gene amplification with high specificity,sensitivity and rapidity,which can be applied for disease diagnosis in shrimp aquaculture.The method is performed under isothermal conditions with a set of four specially designed primers that recognize six distinct sequences of the target.In the present study,a set of four specific primers were desi... |
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| Keywords: | shrimp white spot syndrome virus loop-mediated isothermal amplification rapid detection |
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