目录
摘要Ⅱ
关键词Ⅱ
AbstractⅢ
引言
引言1
文献综述2
1 材料与方法3
1.1 材料 3
1.2 方法 4
1.2.1 金黄色葡萄球菌细菌培养及特性观察4
1.2.1.1培养特性观察4
1.2.1.2革兰染色特性观察4
1.2.1.3生化试验4
1.2.2 金黄色葡萄球菌PCR鉴定5
1.2.3 金黄色葡萄球菌全菌抗原的制备6
1.2.4 实验兔免疫6
1.2.5采血及保存抗血清6
1.2.6 ELISA测定金黄色葡萄球菌抗体效价6
2 结果与分析7
2.1 金黄色葡萄球菌细菌培养及特性观察7
2.1.1生长特性8
2.1.2革兰染色特性8
2.1.3生化特性8
2.2 金黄色葡萄球菌PCR鉴定9
2.3 金黄色葡萄球菌全菌抗原制备9
2.4 实验兔免疫10
2.5 抗体效价测定10
3 讨论 10
致谢11
参考文献12
金黄色葡萄球菌多克隆抗体的制备
摘要
金黄色葡萄球菌可导致人与动物多种疾病，也是引起人类细菌性食物中毒的重要病原，因此在食品卫生上是常规病原菌检测项目之一。通过免疫动物获得的多克隆抗体可同时识别和结合多个抗原表位，而且因制备技术简单、来源方便，在免疫学研究中使用广泛。本研究针对实验室保存的金黄色葡萄球菌菌株进行细菌培养及特性观察，制备全菌抗原，对实验兔进行免疫获得抗血清，最后测定金黄色葡萄球菌抗体效价。结果显示，金黄色葡萄球菌在LB琼脂培养基上生长良好，菌落呈光滑奶油状的圆形小凸起状，边缘完整；革兰染色见紫色深染、排列成葡萄串状的球菌，无鞭毛和芽胞；生化反应中，可分解多种糖类物质，产酸不产气。使用1mL 浓度为3×108 cfu/mL的金黄色葡萄球菌全菌抗原对实验兔进行三次皮内多点注射免疫，每次免疫间隔910 d，并在第三次免疫两周后对实验兔心脏采血，收集抗血清，E *51今日免费论文网|www.51jrft.com +Q: ^351916072* 
LISA检测抗体效价为1:3200。本实验成功制备效价较高的金黄色葡萄球菌多克隆抗体，为建立该菌快速检测方法提供材料。
PREPARATION OF POLYCLONAL ANTIBODIES AGAINST STAPHYLOCOCCUS AUREUS
ABSTRACT
Staphylococcus aureus is an important cause of food bacterial poisoning in humans. It can cause a variety of diseases in humans and animals. It is one of the routine pathogen detection items. The polyclonal antibodies obtained by immunizing animals can simultaneously recognize multiple epitopes of antigens. Since the preparation technology of polyclonal antibodies is simple, and the serum source is convenient, so they are widely used in immunological experiments. In this study, one strain of S. aureus was cultured and its characteristics were observed. The whole bacterial antigen was prepared to immunize the experimental rabbits and obtain the antiserum. Finally, the titer of the antibody against this bacterium was determined. The results showed that S.aureus grew well on the LB agar medium, and the colonies were smooth and creamy, with small round ridges and complete edges. Gram staining showed small globular clusters of grapes with deep purple staining, without flagella and buds, which were grampositive cocci. Biochemical reaction indicated that this bacterium could decompose a variety of sugars, with production of acids but not gas.The experimental rabbit was immunized with 1 mL of 3×108 cfu/mL S. aureus antigen by intradermal multipoint injection for three times, with an interval of 9 to 10 days for each immunization. Blood sample was collected from the hearts of the experimental rabbits after the third immunization for two weeks, and the serum was harvested.The antibody titer was 1:3200 by ELISA. The experiment obtained the highefficient polyclonal antibody against S. aureus by using the routine polyclonal antibody preparation procedure, whichh provides a research basis for the establishment of a rapid detection method for this bacterium.

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