摘要
目的:建立基于问号钩端螺旋体(简称钩体)rLipL32/1-LipL21-OmpL1/2融合抗原的ELISAs,并应用于检测钩体病患者血清特异性IgG和IgM。方法:采用显微镜凝集试验(MAT),检测钩体病患者血清及rLipL32/1-LipL21-OmpL1/2兔抗血清对我国问号钩体参考标准株的效价。分别以rLipL32/1-LipL21-OmpL1/2、rLipL32/1、rLipL21和rOmpL1/2为包被抗原,建立检测血清特异性IgM和IgG的ELISAs,并用于检测107份MAT阳性钩体病患者血清标本。结果:MAT结果证实,66%(71/107)患者感染黄疸出血群问号钩体,rLipL32/1-LipL21-OmpL1/2兔抗血清对我国15株问号钩体参考标准株的凝集效价为1∶20～1∶160。rLipL32/1-LipL21-OmpL1/2、rLipL32/1、rLipL21、rOmpL1/2抗原用于检测IgM的ELISA阳性率分别为89.7%、75.7%、85.1%和79.4%,用于检测IgG的ELISA阳性率分别为99.1%、99.1%、94.4%和86.0%。rLipL32/1-LipL21-OmpL1/2-IgM-ELISA检测阳性率高于其它3种检测IgM的ELISAs(P<0.05),rLipL32/1-LipL21-OmpL1/2-IgG-ELISA检测阳性率高于rOmpL1/2-IgG-ELISA(P<0.05),但与rLipL21-IgG-ELISA及rLipL32/1-IgG-ELISA接近(P>0.05)。结论:rLipL32/1-LipL21-OmpL1/2-ELISAs可作为敏感、特异的通用型钩体病血清学早期诊断方法。
Objective： To establish ELISAs based on rLipL32/1-LipL21-OmpL1/2 fusion antigen of Leptospira interrogans for detecting specific IgG and IgM in serum of patients with leptospirosis. Methods. Microscope agglutination test （MAT） was performed to detect serum specimens from leptospirosis patients and to determine titers of rabbbit antiserum agaist rLipL32/ 1-LipL21-OmpL1/2 to reference standard strains of L. interrogans. By using rLipL32/1-LipL21 -OmpL1/2, rLipL32/1, rLipL21 and rOmpL1/2 as the coated antigens, ELISAs for detectiong sepcific serum IgM and IgG were established. The established ELISAs were applied to MAT -positive serum specimens from 107 patients with leptospirosis. Results. The results of MAT confirmed that 66% （71/107） of the patients were infected with L. interrogans serogroup Icterohaemorrhagiae,and the rLipL32/1-LipL21-OmpL1/2 antiserum were able to agglutinate all 15 reference standard L. interrogans strains with 1 ： 20- 1 ： 160 titers. The positive rates of ELISAs using rLipL32/1-LipL21-OmpL1/2, rLipL32/1, rLipL21 or rOmpL1/2 as the antigen were 89.7%,75.7%,85.1% and 79.4% for detecting IgM,respectively,while 99.1%,99.1%, 94.4% and 86.0% for detecting IgG, respectively. The positive detection rate of rLipL32/1 -LipL21-OmpL1/2-IgM-ELISA was higher than those of the other three IgM detection ELISAs （P〈0. 05）. The positive detection rate of rLipL32/1-LipL21-OmpL1/2-IgG-ELISA was higher than that of rOmpL1/2-IgG-ELISA （P〈0. 05）,while there was no significant differnce with that of rLipL21-IgG-ELISA and rLipL32/1-IgG-ELISA （P〉0.05）. Conclusion： The ELISAs using rLipL32/1-LipL21-OmpL1/2 as the antigen can be applied as a sensitive,specific and universal serological method for diagnosis of leptospirosis, rLipL32/1-LipL21-OmpL1/2-IgM-ELISA shows a definite value for early diagnosis of leptospirosis compared with the other ELISAs used in this study.
出处
《浙江大学学报（医学版）》
CAS
CSCD
2008年第6期592-598,共7页
Journal of Zhejiang University(Medical Sciences)
基金
浙江省科技厅国际合作重点项目(2006C24003)
