淋球菌外膜Porin Ⅰ蛋白多价抗血清的制备及其临床应用初探  

Preparation and Clinical Application of Rabbit Antiserium Against Recombinant Outer Membrane Protein Porin Ⅰ of Nesseria Gonorrhoeae

作　　者：岑建萍[1] 程浩[1] 曾凤英[1] 郑磊[1] 卢忠明[1] 

机构地区：[1]浙江大学医学院附属邵逸夫医院皮肤科,浙江杭州310016

出　　处：《中国皮肤性病学杂志》2007年第9期531-533,共3页The Chinese Journal of Dermatovenereology

基　　金：浙江省卫生厅科研基金(编号491030-W10001);浙江省科委科研基金(编号491030-J30017)

摘　　要：目的制备淋球菌外膜GST-PⅠ融合蛋白多价抗血清,并探讨该多价抗血清与临床泌尿生殖道分泌物标本的结合情况。方法重组的淋球菌外膜GST-PⅠ融合蛋白免疫家兔,获得多价抗血清;并用双向免疫扩散法鉴定家兔多价抗血清的效价。家兔多价抗血清与临床泌尿生殖道分泌物标本的结合采用试管凝集法,出现棉絮状沉淀则为阳性;并把同一标本的试管凝集法结果与淋球菌培养结果相比较,观察两者的阳性符合率。同时以生理盐水和非淋菌性尿道炎患者的泌尿生殖道分泌物标本作为对照。结果成功免疫家兔并获得了PI蛋白多价抗血清,经鉴定多价抗血清的效价达1:8;淋球菌培养阳性的35例分泌物标本中,试管凝集法阳性23例,两方法结果符合率为65.71%。而对照组试管凝集法结果均为阴性。结论淋球菌外膜PI蛋白不仅具有一定的免疫原性,而且其免疫家兔产生的多价抗血清与淋病患者的分泌物标本有良好的结合率,特异性高。有望经纯化、标记后得到一种新的简单快速的淋病诊断试剂盒。Objective To explore the preparation and clinical application of rabbit antiserum against recombinant outer membrane protein Porin I （PI） of Nesseria Gonorrhoeae. Methods Antiserum against gonococcal PI was raised in rabbits by immunizing with highly purified recombinant gonococcal GST-PI fusion protein. The titer of antiserum was detected by double agar immunodiffusion with GST-PI fusion protein. Clinical urogenital secretions were both cultured for gonococcals and reacted with antiserum by direct agglutination in the tube. The result of agglutination was judged as positive when precipitation appeared and was compared with that of gonococcal culture. At last the positive coincidental rate of gonococcal culture and direct agglutination were calculated. Normal saline and urogenital secretions from patients with nongonococcal urethritis were served as control group. Results We successfully prepared rabbit antiserum against gonococcal PI by immunizing with recombinant GST-PI fusion protein and the titer of antiseirum detected by doubleagar immunodiffusion was 1 ： 8. Of all the 35 clinical specimens whose gonococcal cultures were positive ,23 cases were also positive by direct agglutination, while 12 cases were negative. Thus the positive coincidental rate of direct agglutination and gonococcal culture was 65.71%. The results of direct agglutination of control groups were all negative. Conclusion The recombinant GST-PI fusion protein has strong immunogenicity and its rabbit antiserum can react with the urogenital secretions from gonorrhoea patients. The high specificity and sensitivity of the direct agglutination utilizing the antiserum indicate that it should be possible to detect the clinical samples by direct agglutination with the antiserum against gonococcal PI.

关 键 词：奈瑟氏球菌 细茵外膜蛋白质类 抗血清 

分 类 号：R378.1[医药卫生—病原生物学]