Institutional Repository of Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences
| Development of a triplex loop-mediated isothermal amplification method for rapid on-site detection of three Vibrio species associated with fish diseases | |
Yu, Lan-ping1,2; Hu, Yong-hua1; Zhang, Xiao-hua3; Sun, Bo-guang1 ; Sun, BG
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| 2013-11-15 | |
| 发表期刊 | AQUACULTURE
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| ISSN | 0044-8486 |
| 卷号 | 414页码:267-273 |
| 文章类型 | Article |
| 摘要 | A triplex loop-mediated isothermal amplification (triLAMP) method was developed to detect three major fish pathogens, i.e., Vibrio harveyi, Vibrio anguillarum and Vibrio alginolyticus. Three sets of four species-specific primers were designed with one or two restriction enzyme sites contained in the inner primers of each set. TriLAMP is performed with a combination of all three sets of primers in a single reaction. A subsequent restriction enzyme analysis is used for discriminating the specific Vibrio species which caused the positive reaction. The specificity of this method was validated against a total of 32 strains representing 19 different bacterial species. The sensitivities of the triLAMP assays were 102 to 103 times higher than those of the traditional PCR assays. Via modified processes of sample preparation and visual inspection of the LAMP products with fluorescent staining, the triLAMP method allowed convenient and sensitive detection of V. harveyi, V. anguillarum and V. alginolyticus in the tissues of experimentally infected fish. These results indicate that the triLAMP method developed in this study is a useful tool for sensitive and rapid on-site detection of Vibrio infection in fish. (C) 2013 Elsevier B. V. All rights reserved.; A triplex loop-mediated isothermal amplification (triLAMP) method was developed to detect three major fish pathogens, i.e., Vibrio harveyi, Vibrio anguillarum and Vibrio alginolyticus. Three sets of four species-specific primers were designed with one or two restriction enzyme sites contained in the inner primers of each set. TriLAMP is performed with a combination of all three sets of primers in a single reaction. A subsequent restriction enzyme analysis is used for discriminating the specific Vibrio species which caused the positive reaction. The specificity of this method was validated against a total of 32 strains representing 19 different bacterial species. The sensitivities of the triLAMP assays were 102 to 103 times higher than those of the traditional PCR assays. Via modified processes of sample preparation and visual inspection of the LAMP products with fluorescent staining, the triLAMP method allowed convenient and sensitive detection of V. harveyi, V. anguillarum and V. alginolyticus in the tissues of experimentally infected fish. These results indicate that the triLAMP method developed in this study is a useful tool for sensitive and rapid on-site detection of Vibrio infection in fish. (C) 2013 Elsevier B. V. All rights reserved. |
| 关键词 | Loop-mediated Isothermal Amplification Vibrio Harveyi Vibrio Anguillarum Vibrio Alginolyticus On-site Detection |
| 学科领域 | Fisheries ; Marine & Freshwater Biology |
| DOI | 10.1016/j.aquaculture.2013.08.016 |
| URL | 查看原文 |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS研究方向 | Fisheries ; Marine & Freshwater Biology |
| WOS类目 | Fisheries ; Marine & Freshwater Biology |
| WOS记录号 | WOS:000326164600034 |
| WOS关键词 | CAUSATIVE AGENT ; HARVEYI ; PATHOGEN ; IDENTIFICATION ; ALGINOLYTICUS ; ANGUILLARUM ; DIAGNOSIS ; SHELLFISH |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 引用统计 | |
| 文献类型 | 期刊论文 |
| 条目标识符 | http://ir.qdio.ac.cn/handle/337002/16559 |
| 专题 | 实验海洋生物学重点实验室 |
| 通讯作者 | Sun, BG |
| 作者单位 | 1.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China 3.Ocean Univ China, Dept Marine Biol, Qingdao 266003, Peoples R China |
| 第一作者单位 | 中国科学院海洋研究所 |
| 推荐引用方式 GB/T 7714 | Yu, Lan-ping,Hu, Yong-hua,Zhang, Xiao-hua,et al. Development of a triplex loop-mediated isothermal amplification method for rapid on-site detection of three Vibrio species associated with fish diseases[J]. AQUACULTURE,2013,414:267-273. |
| APA | Yu, Lan-ping,Hu, Yong-hua,Zhang, Xiao-hua,Sun, Bo-guang,&Sun, BG.(2013).Development of a triplex loop-mediated isothermal amplification method for rapid on-site detection of three Vibrio species associated with fish diseases.AQUACULTURE,414,267-273. |
| MLA | Yu, Lan-ping,et al."Development of a triplex loop-mediated isothermal amplification method for rapid on-site detection of three Vibrio species associated with fish diseases".AQUACULTURE 414(2013):267-273. |
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| Development of a tri（805KB） | 限制开放 | CC BY-NC-SA | 浏览 | |||
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