多杀性巴氏杆菌菌体成份分析

将多杀性巴氏杆菌(Pasteurella multocida)P-1059(血清3型)和A-898(荚膜A群)二种菌株分别加热抽提制成粗制菌体抗原或荚膜抗原,再作提纯与分析。结果表明:将抗原粗提物盐析时,沉淀中有效抗原组份随硫酸铵饱和度的增加而增加;经高压液相层析,P-1059菌体抗原分为4个峰,A-898荚膜抗原分为3个峰,二者有效抗原组分均在第一峰中;经SDS-聚丙烯酰胺凝胶电泳,P-1059一峰分出3条蛋白带,相应的分子量约为54.3、45.9和38.3kD;用链霉蛋白酶E、神经氨酸酶、蛋白酶K和高碘酸钠分别处理P-1059一峰和A-898一峰后,经ELISA测定,高碘酸钠对P-1059及A-898一峰有分解作用,说明其相应抗原决定簇可能与糖基有关;ELISA交叉抑制试验结果表明,只有A-893一峰对P-1059一峰有弱抑制,说明P-1059菌体中有微量的荚膜成份,而A-898荚膜抗原中无P-1059菌体抗原成份。

Analysis of the Somatic and Capsular Antigens of Pasteurella Multocida

Crude somatic and capsular antigens were prepared respectively from Pasteurellamultocida P-1059 strain (serotype 3) and A-898 strain (capsular group A) by heat-extraction. The crude extracts(CE) were purified by ammonium sulfate precipitationor high-pressure liquid chromatography (HPLC, G 4000). ELISA, SDS-PAGE andenzyme treatment were used to analyse the biological properties of the purified ma-terials. The results were as follows: (1) Binding activity of the precipitate to anti-body rose with the increase of ammonium sulfate concentration. (2) Four proteinpeaks of P-1059 and three peaks of A-898 were obtained by HPLC. Antigenic reac-tivity was detectable only in the first peak of the both strains. (3) Upon SDS-PAGE, peak 1 of P-1059 showed three protein bands. corresponding to molecular wei-ghts of 54300. 45900 and 38300. (4) Purified materials of P-1059 and A-898 weretreated respectively by pronase E, proteinase K, neuramidinase and sodium periodate.After treated with sodium periodate, the two kinds of antigen were not reactive totheir specific antisera. It suggested that the antigenic determinant may be relative tocarbohydrate. (5) The result of cross inhibition by ELISA between the purifiedmaterials of P-1059 and A-898 indicated that the former contained trace ingredientof the latter.