Pantophlet R, Brade L, Dijkshoorn L, Brade H
Division of Medical and Biochemical Microbiology, Research Center Borstel, Center for Medicine and Biosciences, Germany.
J Clin Microbiol. 1998 May;36(5):1245-50. doi: 10.1128/JCM.36.5.1245-1250.1998.
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 Acinetobacter lipopolysaccharide (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 of Acinetobacter 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 of Acinetobacter strains is feasible and thus may provide a simple method for the routine identification of these opportunistic pathogens.
据报道，不动杆菌引发医院获得性感染的频率日益增加。然而，临床实验室仍缺乏能在种水平准确鉴定不动杆菌菌株的简便方法。在本研究中，我们通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳及用超免疫兔血清进行免疫印迹，对44株临床和环境分离株经蛋白酶K消化的全细胞裂解物展开研究，以探讨基于不动杆菌脂多糖（LPS）的O抗原制定血清分型方案的可能性。用所研究的13株热灭活分离株免疫兔子获得抗血清，通过蛋白质印迹法以及以蛋白酶K消化的全细胞裂解物和酚水提取的LPS作为抗原的酶免疫测定法对其特性进行鉴定。在这两种测定中，抗血清对同源抗原均表现出高度特异性。此外，仅基于银染凝胶结果来判断不动杆菌LPS属于光滑型还是粗糙型并不可靠。通过蛋白质印迹分析确定，在31株分离株中有11株观察到O抗原反应性，其中大多数属于鲍曼不动杆菌（DNA群2）和未命名的DNA群3。有趣的是，在与用于免疫的菌株不同的DNA群中发现了一些O抗原。结果表明，不动杆菌菌株的O血清分型是可行的，因此可能为这些机会致病菌的常规鉴定提供一种简便方法。
