摘要
目的 克隆和表达日本血吸虫免疫素 Sjp5 0的编码基因 ,初步研究其免疫反应性。方法 根据 EST测序的结果设计引物 ,从含有免疫素基因片段的克隆中扩增得到该编码基因 ,亚克隆入原核表达载体 p GEX4 T- 1中表达 ,然后用 EL ISA方法初步检测重组蛋白的免疫反应性。结果 成功克隆和表达了日本血吸虫 Sjp5 0基因。该重组蛋白用于血吸虫抗体的 EL ISA检测 ,在动物实验中检测特异性和敏感性分别为 94 .12 %和 76 .6 7% ,非治疗组兔体内的抗体在感染后 9- 11周上升到高水平 ,并在感染后 2 1周仍维持在高水平 ,而治疗组兔血清中抗 Sj Fp5 0抗体水平在治疗后 11周迅速下降至感染前抗体水平。用于血吸虫病人血清中的抗体检测的特异性为 98.30 % ,急性病人和慢性病人检测的敏感性分别为 95 .2 0 %和 6 3.2 0 %。结论 获得了 Sjp5 0的重组蛋白 ,该蛋白能被日本血吸虫病人的血清所识别 ,且在动物实验中其抗体水平在药物治疗后 11周降至感染前的水平 ,具有诊断现症感染病人和疗效考核的潜能。
Objective To clone and express the immunoplicilin of Schistosoma japonicum (Sjp50) and explore its immunological response. Methods The specific primers were designed according to the EST sequence, which was used for amplification of the encoding sequence from the cDNA clone containing Sjp50. The gene was subcloned into pGEX4T-1 plasmid and expressed. The rSjp50 was tested for its immunogical response by ELISA. Results The gene of Schistosoma japonicum Sjp50 was cloned and expressed successfully. The immunogenicity and diagnostic potential of rSjp50 were investigated. It was demonstrated by immunoassay in rabbits that the specificity and sensitivity of the test were 94.12% and 76.67% respectively. The antibody titer of infected group went up to high level at week 9-11 post-infection and maintained at high level until week 21 post-infection, while the antibody level in the treated group rapidly decreased to pre-infection level at 11 weeks after treatment. In human, the specificity of the test was 98.30%. For the acute patient, the sensitivity of anti- Sjp50 was 95.20%, while for the chronic patient 63.20%. Conclusion The recombinant protein of Sjp50 was acquired. It could be recognized by the serum from patient with schistosomiasis, and the level of antibodies was decreased to pre-infected level at 11 weeks after treatment in experimental rabbits, which implicates the potential application on evaluation of chemotherapy and detection of active infections.
出处
《中国血吸虫病防治杂志》
CAS
CSCD
2003年第1期7-11,共5页
Chinese Journal of Schistosomiasis Control
基金
国家高新技术研究发展计划课题
TMRC/ NIH/USA P5 0 A13 9461-0 5 S1
