猪链球菌EPF基因的分子检测

猪链球菌EPF基因的分子检测(4500字)
摘  要：本研究针对猪链球菌（Streptococcus suis）重要的毒力因子EPF（extracellular protein factor，胞外蛋白因子）基因，设计特异性引物，以猪链球菌DNA为模板，建立了猪链球菌属特异性分子检测方法PCR（Polymerase Chain Reaction，聚合酶链式反应）法。标准菌株、疑似病例分离株在预计的位置扩增出特异性条带，表明这套引物成功扩增出目的基因序列，但副猪嗜血杆菌、巴氏杆菌、大肠杆菌和沙门氏菌等的DNA都没有扩增出目的基因片段。链球菌 PCR 检测方法的建立有利于猪致病性链球菌的快速检测和流行病学调查。
关键词：猪链球菌；EPF基因；分子检测；PCR

Molecular detection for EPF gene of Streptococcus suis
Abstract: The specificity of swine Streptococcus molecular detection method of PCR which is  based on important virulence factor EPF(extracellular protein factor) gene of Streptococcus suis,through designing specific primer and making template as DNA of Streptococcus suis,was established in this study. specific bands were amplified in the expected location of tandard strains and isolated strains from suspected cases, indicating that this set of primers successfully amplified the target gene sequence ,but DNA of Haemophilus parasuis, Pasteurella, Escherichia coli and Salmonella etc were not amplified specific gene fragment. The PCR assay can be both for diagnostic purposes and in epidemiological and transmission studies of porcine Streptococcus suis.          
Key words: Streptococcus suis; EPF gene; Molecular detection; PCR 
 

目    录
摘  要    1
关键词    1
1  前言    2
1.1 病源    2
1.2 流行病学    2
1.3 主要毒力因子    2
1.4 PCR检测    3
2  材料与方法    3
2.1 菌株来源    3
2.2 主要试剂    3
2.3 PCR引物序列    3
2.4 菌体培养    4
2.5 DNA模板的制备    4
2.6 PCR扩增    4
2.7 特异性试验    5
3  结果与分析    5
3.1 标准菌株PCR    5
3.2 特异性试验    5
3.3 敏感性试验    6
4  结论与讨论    7
4.1 结论    7
4.2 讨论    7
参考文献    7
致    谢    9