摘要
根据副猪嗜血杆菌16S rRNA基因设计了一对引物，通过最佳条件摸索扩增出大小为821bp的特异目的基因片段，建立了快速检测副猪嗜血杆菌的PCR方法。该方法最低检出量达10^-3 ng，且对大肠埃希菌、金黄色葡萄球菌、传染性胸膜肺炎放线杆菌和巴氏杆菌等均无交叉反应。用该PCR方法从门诊送检的病料中检测出4株副猪嗜血杆菌，并对分离株SH0854P的PCR扩增产物进行测序与对比分析，其与已发表的GenBank中的相关菌株的同源性为97．3％～100％。
The PCR assay was established for the rapid detection of Haemophilus parasuis with a pair of primers designed according to the specific conserved sequence of 16 S rRNA gene. The amplified product was a fragment with 821 bp in length by groping the optimum conditions. Simultaneously, specificity and sensitivity assays revealed that there is no cross reaction with Escherichia coli, Staphylococcus, Actinobacillus pleuropneumoniae and Pasteurella. it is specific to Haemophilus parasuis with a minimum detection sensitivity of 10.3 ng. Four strains of Haemophilus parasuis were isolated from clinical department,and then the sequence of SH0854P was compared. The results showed that the homology with some published strains was high to 97.3%-100%. The PCR assay is specific, rapid, stable, sensitive and efficient.
出处
《动物医学进展》
CSCD
北大核心
2009年第1期9-12,共4页
Progress In Veterinary Medicine
基金
上海市科委2007年重大科技攻关项目(07dz19508)
