Abstract: Objective To compare the phenol water method and the iTron kit in extracting lipopolysaccharides(LPS) from Brucella. Methods LPS were extracted from B. melitensis strain 16M using phenol water method and kit method. The purity and structure of LPS was compared by SDS-gel electrophoresis and silver stain. The activity of the extracted LPS was detected by limulus amebocyte lysate agglutination test, and the characteristics of the two methods were compared. Results LPS extracted from Brucella by both methods had a high purity. There was no significant difference in activity of LPS extracted by phenol water method and the iTron kit (t'=1.270, P=0.332), which was (4.926±0.051) and (5.015±0.037) EU/ng, respectively. LPS extracted by phenol water method might lose some LPS at 17×10³ and 26×10³, while using the iTron kit we can get intact LPS in a convenient and safe way. Conclusion The iTron kit has advantages in extracting LPS from Brucella compared with the phenol water method.

Key words: Brucella, Lipopolysaccharides, Phenol water method, iTron kit

摘要： 目的 比较酚水法和试剂盒法提取布鲁氏菌脂多糖的效果。方法 用酚水法和试剂盒法分别提取羊种布鲁氏菌标准株16M的脂多糖，用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）和银染方法比较脂多糖纯度和结构的差异。用ELISA法定量测定脂多糖浓度，稀释至相同浓度后通过鲎试剂凝集试验检测并比较脂多糖的凝集活性，并从操作过程等方面对两种方法进行比较。用SAS 9.3软件对所得数据进行统计分析，两组均数之间的比较，两组方差齐用Student t检验；若组间方差不齐，则使用调整的t'检验，以α=0.05进行统计学检验。结果 酚水法和试剂盒法提取的脂多糖纯度均较高，二者的鲎试剂凝集活性分别为（4.926±0.051）和（5.015±0.037）EU/ng，差异无统计学意义（t'=1.270，P=0.332）。酚水法提取的脂多糖在17×10³及26×10³处存在缺失，而试剂盒法提取的脂多糖结构更加完整，过程也更加安全方便。结论 试剂盒法提取布鲁氏菌脂多糖比酚水法更具有优势。

关键词: 布鲁氏菌, 脂多糖, 酚水法, 试剂盒法