大鲵致病性嗜水气单胞菌PCR诊断方法的建立  

Development of PCR for detection of pathogenic Aeromonas hydrophila

作　　者：余波[1] 王芳 罗永成 徐景峨[1] 杨莉[1] 史开志[1] 

机构地区：[1]贵州省畜牧兽医研究所,贵州贵阳550005 [2]贵州省生物研究所,贵州贵阳550009 [3]贵州省水产技术推广站,贵州贵阳550001

出　　处：《广东农业科学》2013年第18期133-135,共3页Guangdong Agricultural Sciences

基　　金：贵州省科技厅农业攻关项目(黔科合NZ字[2012]3023号);贵州省畜禽健康养殖技术创新能力建设项目(黔科合院所创能[2010]4004)

摘　　要：根据GenBank中致病性嗜水气单胞菌气溶素基因(hlyA)序列,设计1对特异性引物,通过对PCR反应条件进行优化,建立了检测大鲵致病性嗜水气单胞菌的PCR诊断方法。结果表明:扩增的阳性条带约为600 bp,特异性、敏感性结果显示,该PCR方法对致病性嗜水气单胞菌DNA的最低检测量为0.4 ng/L,而非致病性嗜水气单胞菌、大肠杆菌、黄杆菌、弗氏柠檬酸杆菌的扩增结果均为阴性。对123份大鲵病料进行检测,结果建立的PCR方法检测结果与细菌学和生化检验结果符合率为97.6%,表明该PCR方法能够对大鲵致病性嗜水气单胞菌样品进行快捷、灵敏、准确的检测。According to the gene sequences of hlyA gene in GenBank, one pairs of specific primer was designed for amplifying the specific fragments of hlyA gene. After optimization of annealing temperature and primers concentrations, PCR was established for simultaneous detection of the hlyA gene. The specific band of 600 bp was amplified. The sensitivity and specificity tests showed that the PCR was highly sensitive in 0.4 ng/L DNA. No band was amplified from nonpathogenetic A. hydrophila, E. coil, Flavobacterium, Citrobacter freundii by PCR. 123 clinical samples were detected by the PCR, conventional microbiology methods and their coherenee was 97.6%. The results revealed that the established PCR assay was sensitive, specific and it could be used to detect pathogenetic A. hydrophila rapidly.

关 键 词：致病性嗜水气单胞菌 PCR 气溶素基因 

分 类 号：S852.614[农业科学—基础兽医学]