Bossé J T, Johnson R P, Rosendal S
Department of Veterinary Microbiology and Immunology, Ontario Veterinary College, University of Guelph.
Can J Vet Res. 1990 Jun;54(3):320-5.
Capsular polysaccharides (CPS) of serotypes 1, 2, 5 and 7 of Actinobacillus (Haemophilus) pleuropneumoniae were obtained from 18 h culture supernatants by precipitation with hexadecyltrimethyl-ammonium bromide (Cetavlon) followed by extraction with sodium chloride and reprecipitation in ethanol. These crude extracts, and portions purified further by phenol extraction to remove contaminating proteins, were evaluated as antigens for the detection of serotype-specific antibodies to A. pleuropneumoniae in sera from immunized rabbits and swine by an enzyme-linked immunosorbent assay. The crude extracts reacted strongly with homologous antisera, but except for serotype 1 showed considerable cross-reactivity with antisera to other serotypes. Phenol extraction greatly improved the serospecificity of the antigens from serotypes 1, 7 and, to a lesser extent, 5. The serotype 2 CPS antigen showed poor reactivity following phenol extraction, and did not appear as useful for detection of serotype-specific antibodies.
猪胸膜肺炎放线杆菌（嗜血杆菌）1、2、5和7型的荚膜多糖（CPS）是从18小时培养上清液中获得的，方法是先用十六烷基三甲基溴化铵（西曲溴铵）沉淀，然后用氯化钠萃取，再在乙醇中重沉淀。这些粗提物，以及通过苯酚萃取进一步纯化以去除污染蛋白的部分，通过酶联免疫吸附测定法评估为抗原，用于检测免疫兔和猪血清中针对猪胸膜肺炎放线杆菌的血清型特异性抗体。粗提物与同源抗血清反应强烈，但除1型外，与其他血清型的抗血清有相当程度的交叉反应。苯酚萃取大大提高了1、7型以及在较小程度上5型抗原的血清特异性。2型CPS抗原在苯酚萃取后反应性较差，似乎对检测血清型特异性抗体无用。
