摘要
目的以亮氨酸氨肽酶(LAP)作为检测抗原,观察日本血吸虫感染后不同时期动物血清中相应抗体的动态变化特征。方法应用免疫印迹试验(Western blot)鉴定日本血吸虫LAP(SjLAP)重组蛋白的表达,并利用His镍柱纯化目的蛋白。采用酶联免疫吸附试验(ELISA)检测感染前、后不同时期小鼠及猪血清中LAP诱导抗体产生的变化特征。结果重组蛋白SjLAP可与anti-His单克隆抗体反应,并可被日本血吸虫感染后小鼠血清所识别。获得高浓度重组蛋白SjLAP,以该蛋白为抗原,采用ELISA法检测发现感染后4周猪血清和感染后6周小鼠血清抗体水平达到高峰;且随着虫卵大量出现,抗体水平均显著下降(P均<0.01)。结论 SjLAP在感染早期的高表达可能对日本血吸虫病早期诊断具有一定的参考价值。
Objective To investigate the antibody response to leucine aminopeptidase in the different stages of Schistosoma japonicum infection.Methods The expression product of SjLAP was identified by Western blot and further purified by using nickel column.The IgG levels in the response to SjLAP in murine and porcine sera were detected by ELISA at different time points after the infection of S.japonicum.Results SjLAP expressed by E.coli was recognized by anti-his monoclonal antibody and S.japonicum-infected mice sera by Western blot.The results of ELISA showed that IgG responses to SjLAP rose gradually and reached the peak at 4 weeks post-infection for pigs（P1） and 6 weeks post-infection for mice（P2）.With the appearance of a large number of eggs in the tissue,SjLAP-specific IgG levels were significantly down-regulated（P1=0.0004,P2=0.0001）.Conclusion SjLAP originated from the adult worm might become a potential target for early diagnosis of schistosomiasis japonica.
出处
《中国血吸虫病防治杂志》
CAS
CSCD
北大核心
2011年第2期163-167,共5页
Chinese Journal of Schistosomiasis Control
基金
国家重大科技专项(2008ZX10004-011)
江苏省自然科学基金(BK2006228)
关键词
日本血吸虫
亮氨酸氨肽酶
抗体应答
小鼠
Schistosoma japonicum
Leucine aminopeptidase
Antibody response
Mouse
