Qiu Xiao-feng, Xu Han-fei, Guo Zhong-qi, Wang Jiang, Yan Jie
Department of Microbiology and Parasitology, College of Medicine, Zhejiang University, Hangzhou 310058, China.
Zhejiang Da Xue Xue Bao Yi Xue Ban. 2008 Nov;37(6):592-8. doi: 10.3785/j.issn.1008-9292.2008.06.009.
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.
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 detecting specific serum IgM and IgG were established. The established ELISAs were applied to MAT-positive serum specimens from 107 patients with leptospirosis.
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 : 20approximate, equals1 : 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).
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.
构建基于问号钩端螺旋体rLipL32/1-LipL21-OmpL1/2融合抗原的酶联免疫吸附测定（ELISA）方法，用于检测钩端螺旋体病患者血清中的特异性IgG和IgM。
采用显微镜凝集试验（MAT）检测钩端螺旋体病患者的血清标本，并测定兔抗rLipL32/1-LipL21-OmpL1/2抗血清对问号钩端螺旋体参考标准菌株的效价。以rLipL32/1-LipL21-OmpL1/2、rLipL32/1、rLipL21和rOmpL1/2作为包被抗原，建立检测血清特异性IgM和IgG的ELISA方法。将建立的ELISA方法应用于107例MAT阳性的钩端螺旋体病患者血清标本。
MAT结果证实，66%（71/107）的患者感染了出血性黄疸钩端螺旋体血清群，rLipL32/1-LipL21-OmpL1/2抗血清能够以1∶20（约等于1∶160）的效价凝集所有15株问号钩端螺旋体参考标准菌株。以rLipL32/1-LipL21-OmpL1/2、rLipL32/1、rLipL21或rOmpL1/2作为抗原检测IgM的ELISA阳性率分别为89.7%、75.7%、85.1%和79.4%，检测IgG的阳性率分别为99.1%、99.1%、94.4%和86.0%。rLipL32/1-LipL21-OmpL1/2-IgM-ELISA的阳性检测率高于其他三种IgM检测ELISA（P<0.05）。rLipL32/1-LipL21-OmpL1/2-IgG-ELISA的阳性检测率高于rOmpL1/2-IgG-ELISA（P<0.05），而与rLipL21-IgG-ELISA和rLipL32/1-IgG-ELISA相比差异无统计学意义（P>0.05）。
以rLipL32/1-LipL21-OmpL1/2作为抗原的ELISA可作为一种敏感、特异且通用的钩端螺旋体病血清学诊断方法。与本研究中使用的其他ELISA相比，rLipL32/1-LipL21-OmpL1/进行全文翻译。2-IgM-ELISA在钩端螺旋体病早期诊断中具有一定价值。
