两种方法提取布鲁氏菌脂多糖的比较  

Comparison of two methods of extracting lipopolysaccharides from Brucella

作　　者：韩秀瑞 田国忠[1] 姜海[1] 赵鸿雁[1] 赵忠智[2] 朴东日[1] 杨镒如 崔步云[1] 

机构地区：[1]中国疾病预防控制中心传染病预防控制所传染病预防控制国家重点实验室感染性疾病诊治协同创新中心,北京102206 [2]青海省地方病预防控制所,西宁811602 [3]北京理工大学生命学院,北京100081

出　　处：《中国媒介生物学及控制杂志》2017年第5期470-472,共3页Chinese Journal of Vector Biology and Control

基　　金：国家自然科学基金(81271900);新疆维吾尔自治区自然科学基金(2016D01A065)~~

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17×10~3及26×10~3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。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 17×10^3and 26×10^3,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.

关 键 词：布鲁氏菌 脂多糖 酚水法 试剂盒法 

分 类 号：R516.7[医药卫生—内科学]