MenA PS-EA偶联物的制备及其在多糖抗体检测中的应用  

Preparation of MenA PS-EA Conjugate and Its Application in Detection of Capsular Polysaccharide Antibody

作　　者：肖丽君[1] 赵恩山 孔健[3] 蒋先敏[3] 

机构地区：[1]承德医学院免疫微生物教研室,承德067000 [2]承德钢厂医院,承德060003 [3]北京绿竹生物技术有限责任公司,北京101113

出　　处：《中国生物制品学杂志》2008年第7期611-614,共4页Chinese Journal of Biologicals

基　　金：国家科技部创新基金资助项目(03C26211100767);承德市2008年科学技术研究与发展项目(200821033)

摘　　要：目的制备MenA PS-EA偶联物及以MenA PS-EA为包被抗原的A群脑膜炎奈瑟菌多糖IgG抗体的ELISA检测试剂。方法培养A群脑膜炎奈瑟菌,提取多糖(MenA PS),与卵清蛋白(EA)以还原氨基法偶联,Sepharose CL-4B纯化。制备以MenA PS-EA为包被抗原的A群脑膜炎奈瑟菌多糖IgG抗体的ELISA检测试剂,考察试剂的精密性和稳定性,并与血清杀菌力试验(SBA)比较。结果纯化的MenA PS-EA经高效液相色谱检测,纯度为81.50%;以MenA PS-EA为包被抗原的ELISA检测试剂精密性和稳定性良好;将该试剂与SBA法比较,其敏感性为96.91%,特异性为90.00%,一致性为95.83%。两种检测方法之间差异无显著意义。结论以纯化MenA PS-EA作为包被抗原建立的A群脑膜炎奈瑟菌多糖特异性IgG抗体ELISA检测试剂可用于检测多糖IgG抗体,为进一步研究及开发细菌多糖类抗体诊断试剂提供参考。Objective To prepare MenA PS-EA conjugate and develop an ELISA kit for IgG against capsular polysaccharide of Nesseria meningitidis serogroup A using the MenA PS-EA as coating antigen. Methods Capsular polysaccharide MenA PS was extracted from the culture of Nesseria rneningitidis serogroup A and coupled to egg albumin （ EA ） by reductive amination. The prepared MenA PS-EA conjugate was purified by Sepharose CL-4B chromatography and used as the coating antigen for development of an indirect ELISA kit for IgG against capsular polysaccharide of Nesseria meningitidis serogroup A. Evaluate the precision and stability of the developed kit. Compare the detection result by the kit with that by serum bactericidal assay （SBA）. Results The purity of purified MenA PS-EA determined by HPLC reached 81. 50%. The developed ELISA kit showed good precision and stability. The sensitivity and specificity of the developed ELISA kit were 96.91% and 90. 00% respectively. The coincidence rate of detection results by the kit and SBA was 95. 83%. Conclusion The developed ELISA kit using purified MenA PS-EA as coating antigen was suitable for the detection of IgG against capsular polysaccharide of Nesseria meningitidis serogroup A. The study provided a basis for further development of detection kit for bacterial polysaccharide antibody.

关 键 词：A群脑膜炎奈瑟菌 多糖 IgG抗体 卵清蛋白 偶联物 ELISA试剂 

分 类 号：R378.15[医药卫生—病原生物学] R392-33[医药卫生—基础医学]