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.
HAN Xiu-rui, TIAN Guo-zhong, JIANG Hai, ZHAO Hong-yan, ZHAO Zhong-zhi, PIAO Dong-ri, YANG Yi-ru, CUI Bu-yun
. Comparison of two methods of extracting lipopolysaccharides from Brucella[J]. Chinese Journal of Vector Biology and Control, 2017
, 28(5)
: 470
-472
.
DOI: 10.11853/j.issn.1003.8280.2017.05.015
[1] 蒋建新. 细菌内毒素基础与临床[M]. 北京:人民军医出版社, 2004:16-200.
[2] Leong D,Diaz R,Milner K,et al. Some structural and biological properties of Brucella endotoxin[J]. Infect Immun,1970,1(2):174-182.
[3] Rittig MG,Kaufmann A,Robins A,et al. Smooth and rough lipopolysaccharide phenotypes of Brucella induce different intracellular trafficking and cytokine/chemokine release in human monocytes[J]. J Leukoc Biol,2003,74(6):1045-1055.
[4] Adone R,Muscillo M,La Rosa G,et al. Antigenic,immunologic and genetic characterization of rough strains B. abortus RB51, B. melitensis B115 and B. melitensis B18[J]. PLoS One,2011,6(10):e24073.
[5] Lapaque N,Moriyon I,Moreno E,et al. Brucella lipopolysaccharide acts as a virulence factor[J]. Curr Opin Microbiol,2005,8(1):60-66.
[6] 王照娜. 大肠杆菌脂多糖的提取纯化及活性分析[D]. 天津:南开大学,2008.
[7] Kianmehr Z,Soleimanjahi H,Ardestani SK,et al. Influence of Brucella abortus lipopolysaccharide as an adjuvant on the immunogenicity of HPV-16 L1VLP vaccine in mice[J]. Med Microbiol Immunol,2015,204(2):205-213.
[8] Kianmehr Z,Kaboudanian Ardestani S,Soleimanjahi H,et al. Comparison of biological and immunological characterization of lipopolysaccharides from Brucella abortus RB51 and S19[J]. Jundishapur J Microbiol,2015,8(11):e24853.
[9] 成鹰. 大肠杆菌和布鲁氏菌脂多糖刺激条件下巨噬细胞差异表达miRNAs的鉴定及其作用机制[D]. 海口:海南大学,2014.
[10] 康洁. 3种方法提取大肠杆菌(E. coli)脂多糖的比较[J]. 中国农学通报,2010,26(21):12-15.
[11] 袁莎,陈鹏博,陈创夫. 布鲁氏菌LPS对小鼠巨噬细胞中NLRP3炎症小体转录水平的影响[J]. 中国畜牧兽医,2015,42(1):9-16.
[12] 杨莲芬,程尧章,鲁齐发. 不同毒力布鲁氏菌株内毒素生物学活性的比较研究[J]. 中国地方病防治杂志,1988,3(1):26-29.
[13] Zygmunt MS,Jacques I,Bernardet N,et al. Lipopolysaccharide heterogeneity in the atypical group of novel emerging Brucella species[J]. Clin Vaccine Immunol,2012,19(9):1370-1373.
[14] Conde-'lvarez R, Arce-Gorvel V, Iriarte M,et al. The lipopolysaccharide core of Brucella abortus acts as a shield against innate immunity recognition[J]. PLoS Pathog,2012,8(5):e1002675.
