| 鱼类致病杀鲑诺卡氏菌(Nocardia salmonicida)特异性PCR检测方法的建立 |
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| 引用本文： | 夏洪丽,;鲁义善,;汤菊芬,;蔡佳,;汪美,;夏立群. 鱼类致病杀鲑诺卡氏菌(Nocardia salmonicida)特异性PCR检测方法的建立[J]. 广东海洋大学学报, 2014, 0(4): 56-61 |
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| 作者姓名： | 夏洪丽, 鲁义善, 汤菊芬, 蔡佳, 汪美, 夏立群 |
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| 作者单位： | [1]广东海洋大学水产学院; [2]广东省水产经济动物病原生物学及流行病学重点实验室; [3]广东省教育厅水产经济动物病害控制重点实验室,广东湛江524088 |
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| 基金项目： | 农业部公益性行业科研专项（201203085）; 湛江市科技计划项目（2013B01001）; 广东海洋大学引起人才科研启动费（C13454）; 大学生创新训练项目（1056613015） |
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| 摘 要： | 鱼类诺卡氏菌病是全球性的鱼病,杀鲑诺卡氏菌(Nocardia salmonicida)是引起鱼类诺卡氏菌病的主要病原之一。根据杀鲑诺卡氏菌16S-23S转录间隔区(ITS)序列设计引物,建立特异性PCR检测杀鲑诺卡氏菌的方法。结果表明,筛选的PCR引物可特异性地扩增出杀鲑诺卡氏菌的ITS片段,检测灵敏度(DNA浓度)为28.3 pg·μL-1。检测人工感染杀鲑诺卡氏菌的鱼组织,结果显示,该方法可从未分离到病原菌的病鱼组织中检出阳性片段,较传统细菌分离鉴定方法更为高效灵敏。
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| 关 键 词： | 杀鲑诺卡氏菌 转录间隔区 聚合酶链式反应 检测 |
Establishment of a Specific PCR Assay to Detect Fish Pathogenic Nocardia salmonicida |
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| Affiliation: | XIA Hong-li, LU Yi-shan, TANG Ju-fen, CAI Jia, WANG Mei, XIA Li-qun( Fisheries College of Guangdong Ocean University2. Guangdong Provincial Key Laboratory of Pathogenic Biology and Epidemiology for Aquatic Economic Animals3. Key Laboratory of Diseases Controlling for Aquatic Economic Animals of Guangdong Higher Education Institutions, Zhanjiang 524088, China ) |
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| Abstract: | Nocardia salmonicida is one of the main pathogen of fish nocardiosis. To develop a specific PCR assay for detection of N. salmonicida, primers were designed according to genomic sequence of 16S-23S intergenic spacers （ITS）, and two specific primers were selected in present study. Results indicate that the selected primers are very specific for detecting N. salmonicida and show positive to very little amount of DNA at 28.3 pg. The application of detecting N. salmonicida-infected tissues revealed that the PCR assay can detect positive fragments from infected fish in which N. salmonicida wasn＇t successfully isolated, so the PCR assay, which is more efficient and sensitive, is superior to the traditional method in clinical diagnose for fish pathogenic N. salmonicida. |
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| Keywords: | Nocardia salmonicida intergenic spacers （ITS） polymerase chain reaction detection |
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