鸡鲍氏志贺氏菌脂多糖的提取方法研究

Study on the Extraction and Preparation of Lipopolysaccharide from S.boydii Isolated from Chicken

为了提取和纯化鸡鲍氏志贺氏菌脂多糖(LPS),利用1%葡萄糖肉汤培养基大量培养已分离鉴定的鸡鲍氏志贺氏菌,并应用酚水法提取鸡鲍氏志贺氏菌脂多糖。经聚乙二醇20000浓缩,DNA酶及RNA酶酶解,离心分离,冷冻干燥等制得纯化的LPS。对提取物分别进行糖、蛋白质及核酸含量测定,结果显示,所提取纯化的LPS糖含量为27.06 mg/L,蛋白质含量为0.25 g/L,核酸含量为40.20 mg/L,鲎试剂检测具有凝集活性。上述结果表明:该试验方法能有效提取鸡鲍氏志贺氏菌LPS,提取物纯度较高,核酸及蛋白质含量较低,是提取脂多糖的一种简便快捷的方法。

In this paper,the methods for extraction and purification of the lipopolysaccharide（LPS） from S.boydii previously isolated from chicken were investigated.The S.boydii were cultured using 1% glucose and broth pure and then the LPS was extracted with the method of Phenol-water.After concentration by polyethylene glycol 20000,DNA enzyme and RNA enzyme enzymolysis,centrifugation,and freeze drying,the contents of sugar,protein and DNA were determined.The results showed that the extracts have agglutinate activity of TAL and levels of sugear,protein,and nucleic acid are 27.06mg/L,0.25g/L,and 40.20mg/L,respectively.This indicates that an effective method for extracting S.boydii＇s LPS with high purity of sugar,low levels of NCA and protein has been established,which provided a good foundation for further research on the pathogenicity of S.boydii.