Specificity of Rabbit Antisera against Lipopolysaccharide of Acinetobacter
Specificity of Rabbit Antisera against Lipopolysaccharide of Acinetobacter
复制标题
兔抗血清对不动杆菌脂多糖的特异性
DOI：
10.1128/jcm.36.5.1245-1250.1998
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发表时间：
1998
影响因子：
9.4
通讯作者：
H. Brade
中科院分区：
文献类型：
--
作者：
R. Pantophlet;L. Brade;L. Dijkshoorn;H. Brade
摘要不动杆菌属与医院获得性感染的关系日益密切。然而，临床实验室仍然缺乏简单的方法，允许在物种水平上准确鉴定不动杆菌属菌株。在这项研究中，蛋白酶K消化的全细胞裂解物从44个临床和环境菌株进行了研究，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和免疫印迹与超免疫兔血清，以研究开发的基础上的O抗原的不动杆菌脂多糖（LPS）的血清分型方案的可能性。用13株热灭活菌株免疫家兔，获得抗血清，用蛋白酶K消化的全细胞裂解物和酚水提取的LPS作为抗原，通过Western印迹和酶免疫测定进行鉴定。在这两种测定中，抗血清显示出对同源抗原的高度特异性。此外，仅根据银染凝胶所得结果将不动杆菌LPS分为光滑或粗糙表型是不可靠的。O-抗原反应，通过Western印迹分析确定，观察到11的31株，其中大部分属于鲍曼不动杆菌（DNA组2）和未命名的DNA组3。有趣的是，在与用于免疫的菌株不同的DNA组中发现了一些O抗原。结果表明，O型不动杆菌属菌株的血清学分型是可行的，从而为这些条件致病菌的常规鉴定提供了一种简单的方法。
ABSTRACT Acinetobacter has been reported to be involved in hospital-acquired infections with increasing frequency. However, clinical laboratories still lack simple methods that allow the accurate identification of Acinetobacter strains at the species level. For this study, proteinase K-digested whole-cell lysates from 44 clinical and environmental isolates were investigated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting with hyperimmune rabbit sera to examine the possibility of developing a serotyping scheme based on the O antigen of Acinetobacterlipopolysaccharide (LPS). The antisera, obtained by immunization of rabbits with 13 of the heat-killed isolates investigated, were characterized by Western blotting and enzyme immunoassay by using proteinase K-digested whole-cell lysates and phenol-water-extracted LPS as antigens. In both assays, the antisera were shown to be highly specific for the homologous antigen. In addition, assignment ofAcinetobacter LPS to the smooth or the rough phenotype was shown not to be reliable when it was based only on the results obtained with silver-stained gels. O-antigen reactivity, determined by Western blot analysis, was observed with 11 of the 31 isolates, most of which belonged to the species Acinetobacter baumannii (DNA group 2) and the unnamed DNA group 3. Interestingly, some O antigens were found in a DNA group different from that of the strain used for immunization. The results indicate that O serotyping ofAcinetobacter strains is feasible and thus may provide a simple method for the routine identification of these opportunistic pathogens.