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.
出处
《中国比较医学杂志》
CAS
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)
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)
Mice
Diagnosis
Specific antigen
Two dimension electrophoresis（2DE）
Western blotting
Enzyme-linked immunosorbent assay （ELISA）
