摘要
根据基因库中嗜水气单胞菌16S rRNA基因序列,设计并合成一对特异性引物,用PCR方法对从病死鲢鱼体内分离到的1株嗜水气单胞菌进行扩增,并对PCR反应条件进行优化,同时测试了该方法的特异性和敏感性。特异性试验结果显示,该引物能扩增出680bp的嗜水气单胞菌特异基因片段,与鳗弧菌、溶藻弧菌、副溶血弧菌、温和气单胞菌、迟缓爱德华氏菌鱼、回爱德华氏菌及海豚链球菌无交叉反应;敏感性试验结果显示,该方法最低检测量为10pg嗜水气单胞菌基因总DNA。表明该实验所建立的PCR检测方法可用于鲢鱼嗜水气单胞菌的快速诊断,对有效治疗和控制鱼类嗜水气单胞菌病的流行具有重要意义。
A pair of specific primers was designed and synthesized according to 16S RNA gene sequences of Aeromonas hydrophila from GenBank. The specific DNA fragment of an Aeromonas hydrophila strain separated from dead disease hypophthalmiehthys molitrix was amplified by using PCR. The PCR reaction condition was optimized and the specificity and sensitivity of this PCR method were tested. The results of specificity test indicated that this primer could amplify 680 bp specific DNA fragment from Aeromonas hydrophila and it had no eross-reac tivity with Vibrio anguillarum , Vibrio alginolyticus , Vibrio parahaemolyticus , Aeromonas sobria , Edwardsiella tarde, Edwardsiella ictaluli and Streptococcus iniae. The sensitivity test indicated that 10pg DNA of Aeromonas hydrophila could be detected with this method. Therefore, the PCR method established in this study could be used to detect Aeromonas hydrophila of hypophthalmichthys molitrix rapidly, and it had important significance on the effective diagnosis and control of Aeromonas hydrophila.
出处
《广西农业科学》
CAS
CSCD
2008年第5期681-684,共4页
Guangxi Agricultural Sciences
基金
广西科技计划项目(科技基0639036科技基0731036桂科合0718007B-34)
