篇名:
放線桿菌胸膜肺炎不活化菌苗之研發Development of an Actinobacillus pleuropneumoniae inactivated vaccine
作者:
張惟茗;林敬覆;吳義興;蕭終融
中文摘要:
為進行放線桿菌胸膜肺炎不活化菌苗之研發,種菌之選殖共測試包括21株第1血清型分離株及分屬第1、2、5及11血清型之4株標準株等菌株共25株,所有分離菌株均先經生化試驗及血清學同定確認。菌株之篩選主要針對ApxI細胞毒素之分泌量及菌落型態進行篩選。ApxI細胞毒素之檢測則以酵素免疫斑點法(使用抗ApxI毒素單源抗體檢測)及溶血力價測定等兩種方式進行。在25株菌株中,21株為平滑型菌落,2株為粗糙型,2株為黏液型。以酵素免疫斑點法檢測菌株之ApxI毒素分泌,結果力價之分布由陰性至1:256,差異相當大。菌株溶血力價之分布則由陰性至1:32。不同菌株在酵素免疫斑點法力價或溶血力價與培養於血液培養基所產生之溶血圈大小一致。最後選殖之種菌為第1血清型分離株,菌落為黏液型,酵素免疫斑點法力價及溶血力價分別為1:256及1:32。以豬胸膜肺炎放線桿菌第1血清型種菌進行大量培養所得之菌液,再利用硫酸銨濃縮後檢測其毒素含量減少低於10%,但是毒素活性卻大幅降低,減少超過90%。將菌液不活化後添加商品化油質佐劑之試製菌苗所進行之特性試驗,純粹試驗、無菌試驗及安全試驗等之結果均符合動物用藥品檢驗標準之規定。試製菌苗所進行之效力試驗,計算試製菌苗之防禦指數為1.41,高於標準。
英文摘要:
A selection of a seed strain for development of Actinobacillus pleuropneumoniae (Ap) vaccine was made from 4 type strains and 21 field strains. Among 25 strains, 21 strains were smooth type, 2 were rough type and 2 were mucus type according colony morphology. The criteria of the selection were based on the secretion of ApxI toxin, since it was one of the major factors of pathogenesis, and on colony morphology. The detection of ApxI toxin was made by immnoblotting test and hemolytic test. The distribution of titer by immnoblotting test and hemolytic test were from negative to 1:256 and from negative to 1:32, respectively. It was coordinated with the hemolytic zone appeared in blood agar cultured. The seed strain selected was a field isolate in 1997 with titer of 1:256 and 1:32 tested by above tests.The cytotoxin contained in culture supernatant of Actinobacillus pleuropneumoniae serotype 1 was concentrated by salting out of ammonium sulfate. The amounts of cytotoxin were lost less than 10% after concentration process when measured by immuno-blotting assay. However, the biological activity of cytotoxin was decreased greatly for more then 90% when detected by hemolytic test. The physical properties, purity, sterility, and safety of prepared inactivated vaccine with addition of a commercialized oil adjuvant were all fit the national corresponding regulation. The evaluation of protective efficacy of prepared bacterin was done by mice protection test. The protective index was 1.41 and the inactivated vaccine was proved to be effective.
備註:
年份:西元 2003 年
期別:第 39 期
詳細內容:
胸膜肺炎放線桿菌(Actinobacillus pleuropneumoniae, Ap) 屬一種溶血性革蘭氏陰性菌,感染後可引起豬出血性纖維素性胸膜肺炎。台灣在1976年首次報告以來 [2],目前已蔓延全省,發生率極高。1981年在生長肥育豬調查中,發現以本病之感染為最主要 [3]。而本菌血清型的複雜和其它細菌、病毒的混合感染使得本病更難控制,也造成極大之經濟損失,因此本病的控制亟待解決。抗生素的使用雖可延遲本病的發生和降低感染豬的死亡率,但一旦停藥,本病將再度爆發。因此本病的防治應以疫苗免疫為主。但一般不活化菌苗往往僅能減少豬群的死亡率,而不能阻止慢性感染 [4]。由於在Ap的致病機制中,毒素不但扮演著重要的腳色同時也是重要的保護抗原,實驗顯示豬免疫兩次含Apx 細胞毒素及transferrin-binding proteins的次單位疫苗,能有效保護豬耐過Ap的攻擊及減少肺部病變 [8] 。因此本實驗之目的即在於研發添加重要毒素的菌苗,希望對於本病之防治能有所助益。
附件下載:
附件名稱:
放線桿菌胸膜肺炎
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