钩端螺旋体重组鞭毛蛋白与钩体病患者血清学检测  

Serological testing of leptospriosis patients with recombinant Leptospira interrogansflagellar filaments proteins

作　　者：于洋[1] 张湘燕[2] 张怡轩[1] 秦金红[2] 赵蔚[2] 

机构地区：[1]沈阳药科大学生命科学与生物制药学院,沈阳110016 [2]上海交通大学医学院病原生物学教研室,上海200025

出　　处：《中国人兽共患病学报》2012年第5期438-441,共4页Chinese Journal of Zoonoses

基　　金：国家高技术研究发展计划(863)项目(2006AA02Z176);国家自然科学基金项目钩端螺旋体比较蛋白质组学研究(3077011)联合资助

摘　　要：目的钩端螺旋体鞭毛蛋白FlaB1、FlaB2、FlaB3和FlaB4与钩体病患者血清反应的研究。方法根据钩端螺旋体黄疸出血群赖型赖株(56601)的基因组序列,对4个编码鞭毛丝状体蛋白基因LA2017、LA2019、LA2417、LA2418进行克隆表达,并纯化得到的重组蛋白,进一步制备多克隆抗体。Western Blot检测这4个蛋白在体外培养钩体中的表达情况,ELISA检测在钩体病患者血清中抗体的表达情况。结果 FlaB1、FlaB2、FlaB3和FlaB4均在体外培养的钩端螺旋体56601株中表达;FlaB1、FlaB2和FlaB4的相应抗体在钩体病患者血清中检测出、FlaB1和FlaB2的相应抗体在其它致病性钩体感染患者血清中测出、FlaB1和FlaB4的相应抗体在疫苗免疫正常人血清测出,且均与正常对照具有显著性差异。结论 FlaB1、FlaB2和FlaB4可能与钩体的致病有一定的关系,FlaB1和FlaB4,特别是FlaB1可作为疫苗候选蛋白或诊断蛋白,值得进一步研究。The aim of this paper was to evaluate the expression of 4 flagellar filament proteins of L. interrogans sero- group Icterohaemorrhagiae serovar Lai （56601） and the reactivity with the sera of leptospirosis patients. These 4 flagellar fila- ment proteins of L. interrogans strain 56601 were expressed with prokaryotic vector （pET32） separately. The recombinant proteins were purified and immunized to rabbits to generate polyclonal antibodies. Western Blot showed that these proteins could express in the L. interrogans strain 56601. Enzyme-linked immunosorbent assay demonstrated that FlaB1, FlaB2 and FlaB4 could react with the sera of the patients infected with L. interrogans strain 56601, FlaB1 and FlaB2 could react with the sera of the patient infected with other leptospires and FlaB1 and FlaB4 could reacted with the sera of the vaccinated people. In conclusion, FlaB1 and FlaB4, especially FlaB1 could be used as a vaccine candidate as well as diagnostic protein.

关 键 词：钩端螺旋体 鞭毛蛋白 钩体感染患者血清 

分 类 号：R377.5[医药卫生—病原生物学]