日本血吸虫重组信号蛋白14-3-3(rSj14-3-3)及其单克隆抗体用于诊断的价值

目的探讨纯化的日本血吸虫重组信号蛋白14-3-3(rSj14-3-3)及其相应单克隆抗体对日本血吸虫病的诊断价值。方法利用纯化的rSj14-3-3抗原和可溶性虫卵抗原(SEA)间接ELISA方法检测急、慢性日本血吸虫病患者血清中特异性抗体;以纯化的抗rSj14-3-3单克隆抗体包板,以兔抗rSj14-3-3多抗和酶标羊抗兔多抗为检测系统的酶标夹心法检测患者血清中的循环抗原。结果用rSj14-3-3检测急、慢性血吸虫病患者血清中特异性抗体的阳性率分别为100%和933%,用抗rSj14-3-3单克隆抗体检测循环抗原的阳性率分别为100%和956%,两者联合检测的总阳性率分别为100%和978%;经治疗后2、4、6、12个月,抗体转阴率分别为387%、548%、75%、90%,血清循环抗原转阴率分别为177%、419%、55%、85%,两者联合检测的总转阴率分别为452%、63%、85%、90%;对正常人血清的检测未见假阳性反应,与华支睾吸虫病和钩虫病患者血清出现轻微的交叉反应;用SEA检测抗体的阳性率分别为978%和956%,与正常人及华支睾吸虫病和钩虫病患者血清均有不同程度的交叉反应,治疗后2、4、6、12个月阴转率分别为32%、65%、125%、15%。结论rSj14-3-3检测急慢性血吸虫病患者血清中的特异性抗体和利用抗rSj14-3-3单克隆抗体检测循环抗原具有高度的特异性和敏感性,抗原和单抗可规模化

Evaluation of the diagnostic values of the recombinant signal protein 14-3-3 of Schistosoma japonicum and its monoclonal antibodies

To evaluate the diagnostic value of the recombinant signal protein 14 3 3 of Schistosoma japonicum (rSj14 3 3) and its monoclonal antibodies, specific antibodies to rSj14 3 3 in sera from patients with acute and chronic schistomiasis were detected with the purified rSj14 3 3 antigen and the soluble egg antigen (SEA) by indirect ELISA.Simultaneously, the circulating antigens were detected with anti Sj14 3 3 monoclonal antibody by P M ELISA. The results showed that the positive rates of anti Sj14 3 3 antibodies in sera from acute and chronic cases of schistosomiasis were 100% and 93.3% ,and those of the circulating antigens in both acute and chronic cases were 100% and 95.6% respectively. In case of the combined detection of both antibody and antigen, the total detection rates were found to be 100% and 97.8%. After treatment for 2, 4, 6 and 12 months, the rates of converting to negative results for antibodies were38.7%, 54.8%, 75% and 90%, and those for the circulating antigens were 17.7%, 41.9%, 55% and 85% respectively.. The converting rates to negative results in case of the combined detection of both antibodies and antigens were even more significant, appearing to be 45.2%, 63%, 85% and90% respectively, In sere of patients after effective treatment. No false result was observed in normal sera, but certain cross reactions were observed in individual cases of clonorchiasis and hookworm infection. Meanwhile, the detection rates of antibodies in acute and chronic cases with SEG antigen were 97.8% and 95.6% respectively, and it showed certain degrees of cross reactions with sera of normal individuals and patients with clonorchiasis and hockworm infections. After treatment for 2, 4,6 and 12 months, 3.2%, 6.5%, 12.5% and 15% of patients turned to be negative results. It is concluded that the method of detecting specific antibodies by using the rSj14 2 3 signal protein as antigen or by using its monoclonal antibodies to detect circulating antigens possesses high degrees of specificity and sensitivity, and can be used as a convenient and practicable method for the diagnosis of schistosomiasis.