《矛尾复虾虎鱼病原哈氏弧菌的鉴定及特异性检测方法的研究》-毕业论文设计（学术).docVIP

矛尾复虾虎鱼病原哈氏弧菌的鉴定及特异性检测方法的研究 摘 要：2009年8至9月间，江苏赣榆县多家虾蟹养殖塘混养的矛尾复虾虎鱼大量死亡，主要病变特征为体表出血、肌肉溃疡，从矛尾复虾虎鱼深层溃烂组织及肝脏中分离出优势菌，通过感染实验表明了该菌对矛尾复虾虎鱼有很强的致病性，当分离菌浓度在106至108可引起矛尾复虾虎鱼全部死亡。对分离菌进行相关的形态及理化特性maximum parsimony，MPPCR检测方法。在PCR扩增中，gyrB基因引物A2B3扩增目的片段大小为为363bp，该方法最低能检测0.0046875ng/μL哈氏弧菌模板DNA；toxR基因引物vh-tox-F、vh-tox-R扩增目的片段大小为为390bp，该方法最低能检测0.75ng/μL哈氏弧菌模板DNA。基于哈氏弧菌的gyrB基因和toxR基因建立的PCR检测技术在对由哈氏弧菌所引起的流行病的调查具有重要意义。 关键词：矛尾复虾虎鱼；哈氏弧菌；PCR；检测 毕业设计（论文）外文摘要 Identification and specific detection method of pathogenic Vibrio harveyi from Synechogobius hasta Abstract: In August to September 2009, outbreaks of mass mortality of pond-cultured Synechogobius hasta have occurred in some ponds of Ganyu county, Jiangsu province, and gross signs of disease exhibited haemorrhages at the body surface, ulcer of muscle. A dominate bacteria was isolated from the samples of the deep ulcerate tissues and the liver, after challenge test which proved it’s high pathogenicity to Synechogobius hasta, and all inoculated fish died post-injection with 0.1mL live cells (106to108CFUmL-1) per fish. The biological characteristics were examined, including morphological characteristics, physiological and biochemical characteristics, the 16S rRNA and gyrB genes were sequenced and compared with that of the correlative strains, the MP phylogenetic trees based on 16S rRNA and gyrB genes were constructed using maximum parsimony (MP) infected with Vibrio harveyi. The results showed that it was Vibrio harveyi which caused a large number of complex death of Synechogobius hasta. Then a rapid and accurate polymerase chain reaction (PCR) that will detect V.harveyi were established using two pairs of specific primers designed by gyrB gene and toxR gene as well as the sensitivity of the test and the last for seafood inspection. The PCR could amplify 363 bp gene fragment from chromosomal DNA of V.harveyi based on gyrB, and detect purified chromosomal DNA at a level of as few as 0.0046875ng/μL. The PCR could amplify 390bp gene fragmen