中国人兽共患病学报2011，Vol.27Issue(6)：484-488,5.
基于2-DE和蛋白质组技术筛选的日本血吸虫鸟氨酸氨基转移酶的表达及其诊断应用
The two-dimensional electrophoresis- and proteomics-based screening and expression of ornithine aminotransferase from Schistosoma japonicum (SjOAT) for the diagnosis of schistosomiasis
摘要
Abstract
In order to clone and express ornithine aminotransferase of Schistosoma japonicum (SjOAT) that screened by proteomic techniques and to access its potential value in the immunodiagnosis of S. japonica, SjOAT was amplified by PCR and ligated to vector pET28a(+). After the recombinant plasmid was transformed into E. coli BL21, the expression was induced by IPTG. The fusion protein was purified by Ni2+-NTA chromatography and identified by SDS-PAGE. Western blotting confirmed the immunoreactions of SjOAT with the positive sera of schistosomiasis patients. SjOAT and soluble egg antigen (SEA) were further applied in the diagnosis of human Schistosomiasis japonica using ELISA. The positivity rates were 90.4 ％ in rSjOAT-ELISA and 94.2％ in SEA-ELISA in 52 cases of acute schistosomiasis. The positive rates in 81 cases of chronic schistosomiasis were 86.4 ％ in rSjOAT-ELISA and 82.7 ％ in SEA-ELISA. Statistical analysis revealed no significant difference in sensitivity (P＞0.05) between rSjOAT and SEA. The false positive reaction rates were found to be 1.1％ in rSjOAT-ELISA and 9.6％ in SEA-ELISA when detected in 94 cases of normal control sera. When rSjOAT-ELISA and SEA-ELASA were subjected to the detection in 94 cases of sera from people in endemic area with negative eggs in the feces, the positive reaction rates were found to be 3.2％and 12.8％. Statistical analysis revealed significant difference in specificity between rSjOAT and SEA(P＜0. 05). The rSjOAT-ELISA and SEA-ELISA were used to detect the sera of patients with clonorchiasis, paragonimiasis and hookworm infection. Statistical analysis revealed no significant difference in cross-reactivity between rSjOAT and SEA(P＞0.05). The result indicated SjOAT was promising antigen for serological diagnosis of S. japonica instead of crude SEA.关键词
日本血吸虫/鸟氨酸氨基转移酶/ELISA/诊断Key words
Schistosoma japonicum/ SjOAT/ ELISA/ diagnosis分类
医药卫生引用本文复制引用
胡梅梅,钟政荣,罗庆礼,方婷娜,沈继龙..基于2-DE和蛋白质组技术筛选的日本血吸虫鸟氨酸氨基转移酶的表达及其诊断应用[J].中国人兽共患病学报,2011,27(6):484-488,5.基金项目
国家"863"计划(2006AA02A251)和安徽省自然科学基金资助项目(090413089)联合资助 （2006AA02A251）
