淋病奈瑟菌膜蛋白抗原表位的研究

Investigation of An Epitope on Outer Membrane Protein of Neisseria Gonorrhoeae

目的利用人工合成多肽制备针对淋球菌外膜蛋白PIB抗原表位的特异性多克隆抗体,以期用于淋球菌的检测,为进一步建立新型淋病快速诊断方法提供研究基础。方法根据PIB抗原表位的氨基酸序列合成一段多肽(LD1),并将其与钥孔血蓝蛋白(KLH)偶联构成完全抗原LD1-KLH,免疫动物,收集抗血清,用ELISA和Western blot检测其特异性。结果ELISA检测表明,LD1-KLH具有免疫原性,免疫动物能刺激机体产生较强的体液免疫反应,兔血清抗体和豚鼠血清抗体效价最高分别达到1:3200和1:25600;抗血清能与淋球菌临床株特异性结合,与葡萄球菌、链球菌、大肠杆菌和真菌等其他微生物之间没有交叉反应;Westernblot结果显示,抗血清能识别淋球菌裂解物,产生特异性条带,相对分子质量(Mr)为37300。结论人工合成多肽具有免疫原性,能诱导机体产生识别淋球菌抗原表位的特异性抗体。

Objective To prepare a polyclonal antibody against an epitope on outer membrane protein PIB of Neisseria gonorrhoeae based on chemically synthesized peptide for establishing a rapid diagnostic test for detecting Neisseria gonorrhoeae. Methods A peptide（ LD1 ） was synthesized corresponding to sequence of the domain containing the epitope. LD1 was coupled to keyhole limpet hemocyanin （KLH） and the LD1-KLH conjugate was used for immunization of animals. Antisera produced following immu- nization were detected by ELISA and Western blot. Results Both the antisera of rabbit and guinea pig reacted with LD1-KLH, and the titer around 1：3 200 and 1：25 600 respectively. The antisera were able to react with Neisseria gonorrhoeae but did not react with other microorganism such as Staphylococcus, Streptococcus, Eseheriehia eoli and fungus. Western blot displayed a specifically Mr （molecular mass relatively）37 300 reactive band in gonococcal whole-cell lysate when incubated in antisera. Conclusion The synthetic peptide has the ability to induce immune response against epitope on gonococcal PIB.