Rapid,sensitive detection of Vibrio anguillarum using loop-mediated isothermal amplification
文献类型：CNKI期刊论文
| 作者 | Gao HW(高宏伟); Li FH(李富花); Zhang XJ(张晓军); Wang B(王兵); Xiang JH(相建海)
|
| 发表日期 | 2010-02-15 |
| 出处 | Chinese Journal of Oceanology and Limnology
 |
| 关键词 | Loop-Mediated Isothermal Amplification (LAMP) detection assay empA gene Vibrio anguillarum |
| 英文摘要 | Vibrio anguillarum is an important bacterial pathogen of aquatic organisms and a significant problem in aquatic farming.The rapid detection and identification of V.anguillarum,and other pathogens that infect marine organisms,is crucial to effective disease management.In this study,we developed a loop-mediated amplification (LAMP) assay to detect V.anguillarum in an hour in a single tube without the need for thermal cycling.Conserved regions of the metalloproteinase (empA) gene of V.anguillarum served as the targets for primer design.A fragment of the empA gene was amplified at 65°C in the presence of the primer mixture and Bst DNA polymerase.In the optimized LAMP assay,6.7 pg of V.anguillarum DNA could be detected.Six strains of V.anguillarum and 17 strains of non-V.anguillarum bacteria were used in this study to evaluate the species specificity of the primers.The six V.anguillarum strains gave a positive result in the LAMP assay.This method was also validated in V.anguillarum-infected fish.This LAMP method is more sensitive than PCR in the detection of V.anguillarum and shows good species specificity.The LAMP assay is therefore an effective method for the quick detection of V.anguillarum both in the laboratory and in the field. |
| 文献子类 | CNKI期刊论文 |
| 资助机构 | Supported by the National Basic Research Program of China (973 Program) (No. 2006CB101804) ; the General Administration of Quality Supervision,Inspection and Quarantine of the People's Republic of China (No. 2007IK167) ; National Department Public Benefit Research Foundation (No. 200803012) |
| 卷 | v.28期:01页:64-68 |
| 语种 | 英文; |
| 分类号 | S941 |
| ISSN号 | 0254-4059 |
| 源URL | [http://ir.qdio.ac.cn/handle/337002/192262]  |
| 专题 | 中国科学院海洋研究所 |
| 作者单位 | 1.InstituteofOceanology,ChineseAcademyofSciences 2.ShandongEntry-ExitInspectionandQuarantineBureauofthePeople'sRepublicofChina |
| 推荐引用方式 GB/T 7714 | Gao HW,Li FH,Zhang XJ,et al. Rapid,sensitive detection of Vibrio anguillarum using loop-mediated isothermal amplification. 2010. |
入库方式： OAI收割
来源：海洋研究所
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。