肺炎克雷伯氏菌特异性抗原的筛选与鉴定  

Identification of Specific Diagnostic Antigen for Klebsiella pneumoniae

作　　者：李挺[1] 张丽芳[1] 刘星[1] 刘先菊[1] 刘江宁[1] 李万波[1] 林树柱[1] 

机构地区：[1]北京协和医学院中国医学科学院实验动物研究所,国家中医药管理局人类疾病动物模型三级实验室,北京100021

出　　处：《中国比较医学杂志》2010年第7期21-26,共6页Chinese Journal of Comparative Medicine

基　　金：中央级公益性科研院所基本科研业务费专项资金(DWS200705)

摘　　要：目的寻找肺炎克雷伯氏菌(Klebsiella pneumoniae)血清学检测用特异性抗原。方法双向电泳分离K.pneumoniae总蛋白,通过免疫印迹(Western blotting)与常见病原菌的多抗反应筛选特异性抗原蛋白,原核表达该蛋白并用ELISA法验证。结果获得K.pneumoniae的双向电泳图谱。寻找Western blotting中与K.pneumoniae自身多抗反应而不与与其它病原菌多抗反应的蛋白,质谱鉴定为酸性磷酸酶(Acid phosphatase,GI:238894261)。经表达纯化并以酶联免疫吸附试验(ELISA)验证,证明该蛋白作为包被抗原的灵敏度高,特异性强。结论 K.pneumoniae酸性磷酸酶是适用于该菌感染检测特异性抗原蛋白。Objective To discover novel antigens of Klebsiella pneumoniae for establishing serological assays. Methods Total protein of K. pneumoniae was separated by two-dimension electrophoresis（2DE）,reacting with polyclonal antibodies of common pathogenic bacteria by western blotting. The specific antigen was selected,expressed in E. coli and then evaluated by ELISA. Results 2DE map of K. pneumoniae was obtained. The protein spot that reacted to pAbs of K. pneumoniae but unreacted to other pAbs in Western blotting was identified by mass spectrometry to be acid phosphatase （GI：238894261）. The protein was subsequently expressed and purified. ELISA results had certified its high specificity and sensitivity. Conclusion Acid phosphatase of K. pneumoniae is the novel protein antigen for specific diagnosis of its infection.

关 键 词：小鼠 检测 特异性抗原 双向电泳 免疫印迹 酶联免疫吸附试验(ELISA) 

分 类 号：Q939.91[生物学—微生物学]