【摘要】 从新疆分离的奶牛乳腺炎金黄色葡萄球菌,经TH-16S系统鉴定,选取鉴定结果为733330的优势生化株。毒力测定试验表明,1×108CFU/只的接种量会使小白鼠在24 h内死亡。菌株接种Columbia Broth Modified 37℃摇床100 r/m in培养24 h后,用0.05%戊二醛灭活。灭活培养物经4℃30 m in 17 000 g离心去除细胞碎片,上清用PBS(pH 7.2)4℃透析48 h,PEG20000浓缩10倍,透析,加入终浓度为10 mg/mL的蜂胶-乙醇浸液,作为荚膜多糖免疫原。先分别用荚膜多糖免疫原0.1、0.2、0.4 mL免疫小白鼠,再用组合免疫原(荚膜多糖+109CFU/mL菌体)0.1、0.2、0.4 mL免疫小白鼠。6组小鼠于免疫后14 d连同对照小鼠2组攻击金黄色葡萄球菌2×108CFU/(0.2 mL.只)。结果表明,荚膜多糖免疫原组保护率分别为0、25%和50%,(荚膜多糖+菌体)组保护率分别为25%、75%和100%。更多还原

【Abstract】 733330 strain was identified from Staphylococcus aureus strains isolated from clinical mastitis dairy cow in Xinjiang by the TH-16S system and demonstrated virulent by mouse challenge test.All mice died within 24 hours after being infected with 1×10~8colony-forming units(CFU)per mouse.The strain was cultivated in Columbia Broth Modified(CBM) at 37 ℃ with shaking of 100 r/min for 24 h.Then the bacteria was exterminated by 0.05% glutaraldehyde.Bacterial fragments were discarded after centrifugation with 17 000 g at 4 ℃ for 30 min.The supernate was dialyzed against PBS(pH 7.2,4 ℃,48 h) and concentrated to 10%volume with PEG20000 as crude capsular polysaccharide.The mice were vaccinated by subcutaneous injection separately with 0.1 mL,(0.2 mL) and 0.3 mL per mouse of two sorts of immunogenicity,the capsular polysaccharide and the capsular polysaccharide plus bacterial fragments of TH-16S 733330 S.aureus(final concentration 1×10~9CFU/mL),and were challenged abdominally 14 days later by 2×10~8 CFU per mouse of the S.aureus.The protective ratio of crude capsular polysaccharide plus bacterial fragments was 25%,75% and 100% respectively,which was higher significantly than that of capsular polysaccharide of 0,25%and 50%.更多还原