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Antigenic heterogeneity of outer membrane proteins of nontypable Haemophilus influenzae is a basis for a serotyping system.

机译:不可分型流感嗜血杆菌外膜蛋白的抗原异质性是血清分型系统的基础。

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摘要

A serotyping system for nontypable Haemophilus influenzae (NTHI) was developed by using isolated outer membrane protein (OMP) preparations and rabbit antisera. OMPs of 23 strains were isolated by molecular sieve chromatography of outer membranes in 1.5% sodium deoxycholate buffer. These OMP preparations were relatively free of lipopolysaccharide as determined by silver staining of sodium dodecyl sulfate gels and by dot assay with a monoclonal antibody which is specific for the lipid A of H. influenzae. Three antisera raised to whole organisms were used to serotype 21 of 23 strains with a kinetic enzyme-linked immunosorbent assay. Digestion of OMP preparations with proteinase K removed greater than 90% of the antigenic reactivity, indicating that the system is based on OMP antigens. Marked antigenic heterogeneity of OMPs exists among strains of NTHI. By determining the pattern of serological reactivity of OMPs with the three antisera, isolates were divided into groups based on antigenic differences. Six serotypes were identified. This OMP serotyping system is based on multiple antigenic determinants. Future studies will focus on identifying serotype-specific epitopes to further refine this serological classification scheme for NTHI.
机译:通过使用分离的外膜蛋白(OMP)制剂和兔抗血清,开发了针对不可分型流感嗜血杆菌(NTHI)的血清分型系统。通过在1.5%脱氧胆酸钠缓冲液中进行外膜分子筛色谱分离23株菌株的OMP。这些OMP制剂相对不含脂多糖,这是通过十二烷基硫酸钠硫酸钠凝胶的银染和通过用对流感嗜血杆菌的脂质A具有特异性的单克隆抗体的斑点测定法确定的。通过动力学酶联免疫吸附试验,使用三种针对全生物的抗血清对23株菌株中的21种进行血清分型。用蛋白酶K消化OMP制剂可去除90%以上的抗原反应性,这表明该系统基于OMP抗原。在NTHI菌株之间存在OMPs的明显抗原异质性。通过确定OMP与三种抗血清的血清反应性模式,根据抗原差异将分离物分为几类。确定了六种血清型。该OMP血清分型系统基于多种抗原决定簇。未来的研究将集中于确定血清型特异性表位,以进一步完善NTHI的血清学分类方案。

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