| 胡野,郭宗琪,孙百莉,杨平,严杰.问号钩端螺旋体属特异性LipL41s抗原膜定位及其患者血清抗体检测[J].Chinese journal of Epidemiology,2007,28(8):776-780 |
| 问号钩端螺旋体属特异性LipL41s抗原膜定位及其患者血清抗体检测 |
| Study on the location of membrane and detection of antibody in the sera of genus-specific antigen LipL41s in patients with Leptospira interrogans |
| Received:January 19, 2007 Revised:August 10, 2007 |
| DOI： |
| KeyWord: 钩端螺旋体 基因 LipL41s 重组表达 抗原定位 免疫应答 |
| English Key Word: Leptospira interrogans LipL41s genesv Recombinant expression Antigen location Immune response |
| FundProject:浙江省科技厅国际合作重点资助项目(2006C24003) |
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| Abstract: |
| 目的 确定问号钩端螺旋体(钩体)属特异性脂蛋白抗原LipL41s膜定位及其自然抗体应答情况和抗体类型.方法 采用显微镜凝集试验(MAT)检测四川地区钩体病患者血清标本.IPTG诱导重组原核系统表达目的蛋白rLipL41/1和rLipL41/2,Ni-NTA亲和层析法提纯目的重组表达产物.采用Western blot检测感染不同血清群问号钩体病患者血清与rLipL41s的免疫反应性.采用胶体金免疫电镜技术,对LipL41s进行膜定位.建立基于rLipL41s的ELISA,检测钩体病患者血清中特异性抗体水平及其类型.结果 黄疸出血群是四川地区最主要的优势钩体血清群.不同血清群问号钩体病患者血清均能有效识别LipL41s.LipL41s是位于钩体外膜表面的蛋白分子.156例MAT阳性钩体病患者血清标本中,rLipL41/1和rLipL41/2特异性IgM阳性率分别为84.6%～87.8%和78.2%～83.3%,特异性IgG阳性率分别为69.2%～81.4%和75.0%～80.1%.结论 LipL41s是钩体表面蛋白抗原.自然感染钩体时,LipL41/1和LipL41/2可诱导机体产生IgM和IgG两类血清抗体,且两者之间有广泛的抗原交叉反应.rLipL41/1和rLipL41/2可作为研制通用型钩体基因工程疫苗和检测试剂盒的候选抗原. |
| English Abstract: |
| Objective To determine the location on outer envelope and natural antibody response and types of genus-specific lipoprotein antigen LipL41s in patients with Leptospira interrogans.Methods Microscope agglutination test(MAT)was used to examine leptospirosis patients' serum samples from Sichuan area,China.Ni-NTA affinity chromatography was performed to extract the target recombinant rLipL41/1 and rLipL41/2 products that expressed under inducement of IPTG.Western blot assay was performed to detect the immunoreactivity between the sera from the patients infected with different sero- groups of L.interrogans and rLipL41s.Immune aurosol electron microscopy was selected to locate the position of LipL41s on leptospiral envelope.ELISA based on rLipL41s was established to confirm the level and types of specific antibody.Results L.interrogans serogroup icterohaemorrhagiae remained to be the most dominant leptospiral serogroup in Sichuan area.All the sera from patients infected with different serogroups of L.interrogans could efficiently recognize the LipL41s which were the protein molecular that located on the external surface of leptospiral envelope.In the 156 serum samples from MAT positive leptospirosis patients,the positive rates for rLipL41/1 or rLipL41/2 specific IgM appeared to be 84.6%- 87.8 % and 78.2 %-83.3 %,respectively,while for rLipL41/1 or rLipL41/2 specific IgG they were 69.2 %- 81.4% and 75.0%-80.1%,respectively.Conclusion LipL41s were the leptospiral superficial protein antigen of L.interrogans.Both the LipL41/1 and LipL41/2 could induce serum antibodies IgM and IgG with extensive antigenic-cross reaction during natural infection of L.interrogans in general populations. Hence,rLipL41/1 or rLipL41/2 could be used as the antigen candidate for developing universal genetic engineering vaccine and detection kit. |
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