应用重组信号蛋白14-3-3间接ELISA诊断日本血吸虫病  

Application of indirect ELISA with recombinant signaling protein 14-3-3 of Schistosoma japonicum for the immunodiagnosis of schistosomiasis

作　　者：罗庆礼[1] 王志成[1] 李敏[1] 郑美娟[1] 余轶婧[1] 罗飞[1] 沈继龙[1] 

机构地区：[1]教育部重要遗传病基因资源利用重点实验室(省部共建)

出　　处：《中国人兽共患病学报》2007年第3期231-235,共5页Chinese Journal of Zoonoses

基　　金：国家高技术研究发展计划(863计划)项目(2004AA2Z3570)

摘　　要：目的探讨重组日本血吸虫信号蛋白14-3-3(rSj14-3-3)间接ELISA用于血吸虫病免疫诊断的价值。方法表达并利用纯化的rSj14-3-3与成虫抗原(SjAWA)间接ELISA法分别检测51份急性和49份慢性日本血吸虫病患者和50份正常人血清,以上血清标本同时用SEA致敏的间接血凝试验(SEA-IHA)检测。再以3种方法检测30份华支睾吸虫感染者、24份卫氏并殖吸虫感染者和31份钩虫感染者血清,分析其交叉反应性。结果51份急性和49份慢性血吸虫病患者血清的rSj14-3-3抗体阳性率分别为98.0%和91.8%;SjAWA的检出率分别为96.1%和90.0%;IHA的阳性率分别为98.0%和93.9%。经统计学分析,以上结果无显著性差异(P>0.05)。Sj14-3-3间接ELISA、SjAWA间接ELISA和SEA-IHA对于30份华支睾吸虫感染者的交叉反应性分别为13.3%、20.0%和16.7%,24份卫氏并殖吸虫感染者分别为8.3%、12.5%和12.5%,31份钩虫感染者分别为12.9%、16.1%和12.9%。经统计学分析结果也无显著性差异(P>0.05)。结论rSj14-3-3抗原间接ELISA法诊断日本血吸虫病,具有高度的敏感性和特异性,可用于血吸虫病的免疫诊断。To evaluate the recombinant signaling protein 14-3-3 of Schistosoma japonicum （rSj14-3-3） on the immunodiagnosis of schistosomiasis by indirect ELISA, the expression of rSj14-3-3 in E. coli was induced and purified with affinity chromatography, and the coated antigen concentration and sera dilution were optimized. Sera of patients with acute and chronic schistosomiasis and negative control were tested with rSj14-3-3 and adult worm antigen of Schistosoma japonicum （SjAWA） by indirect ELISA and soluble eggs antigen （SEA）-IHA respectively. It was found that the positive rates of indirect ELISA in 51 cases of acute and 49 of chronic schistosomiasis were 98.0% and 91.8 % using rSj14-3-3, whereas those were 96. 1% and 90.0% using SjAWA. The positive rates of SEA-IHA were 98. 0% and 93.9% in both acute and chronic patients. Two percent of cross-reaction was noted in normal sera tested by rSj14-3-3 antigen and, 6.0% by SjAWA, compared with 2.0 % of false positive in SEA-IHA. There were no statisticaly significant difference of the results among rSj14-3-3, SjAWA, and SEA-IHA （P 〉0. 05）. Meanwhile 13. 3%, 20. 0% and 16. 7% of cross-reaction were observed among rSj14-3-3-ELISA, SjAWA-ELISA and SEA-IHA for detecting the sera of patients with Clonorchis sinensis as well as 8.3% ,12.5% and 12.5% in the cases of Paragonimus westermani infection and 12.9%, 16.1% and 12.9% in the cases of hookworm infection. Therewas no significant difference of cross-reaction in three parasitic infections. The present study showed that the indirect ELISA with rSj14-3-3 antigen had as high sensitivity and specificity as SEA-IHA for diagnosis of schistosomiasis.

关 键 词：日本血吸虫病 14-3-3蛋白 免疫诊断 ELISA间接血凝试验 

分 类 号：R532.1[医药卫生—内科学]