HRP-ASSP用于Dot-ELISA检测日本血吸虫抗体的实验研究

Experimental studies on the detection of Schistosoma japonicum antibodies by Dot-ELISA with HRP-ASSP

目的探讨辣根过氧化物酶标记猪蛔虫可溶性蛋白质(HRP-ASSP)用于Dot-ELISA检测日本血吸虫病患者血清(Sj-S)中抗日本血吸虫抗体的可行性。方法参照改良过碘酸钠法,采用HRP标记ASSP得到HRP-ASSP,经Dot-ELISA观察HRP-ASSP与ASSP免疫兔血清反应,并测定效价;再将HRP-ASSP分别用于直接Dot-ELISA和间接Dot-ELISA检测10份Sj-S,并与检测10份健康人血清(Hp-S)、8份卫氏并殖吸虫病患者血清(P-S)、10份囊尾蚴病患者血清(C-S)和6份旋毛虫病患者血清(T-S)进行比较。结果HRP-ASSP与ASSP免疫兔血清呈阳性反应;直接Dot-ELISA检测Sj-S阳性反应率为100%,Hp-S、P-S、C-S、T-S阳性反应率为20%～50%;间接Dot-ELISA检测Sj-S阳性反应率为100%,其他血清阳性反应率均为0。结论ASSP与日本血吸虫特异性抗体可能具有高亲和力,HRP-ASSP用于间接Dot-ELISA检测Sj-S中抗日本血吸虫抗体具有潜在的应用价值。

Objective To explore the feasibility of detection of Schistosoma japonicum antibodies in the sera of patients with schistosomiasis japonica （Sj-S） by Dot-ELISA with HRP-ASSP obtained by labelling Ascaris suum soluble protein （ASSP） with horseradish peroxidase. Methods The reaction of HRP-ASSP which repowed by labelling ASSP with HRP in accordance with the improved sodium periodate method to the sera of rabbits immunized with ASSP was observed by Dot-ELISA, and its titer was detected. Ten samples of sera of patients with schistosomiasis iaponica were detected by direct Dot-ELISA and indirect Dot-ELISA with HRP-ASSP and compared with 10 samples of sera of healthy persons （Hp-S）, 8 samples of sera of patients with paragonimiasis （P-S）, 10 samples of sera of patients with cysticercosis （C-S） and 6 samples of sera of patients with trichinelliasis （T-S）. Results It was positive that HRP-ASSP reacted with the sera of rabbits immunized with ASSP. The positive reaction rate was 100% in detecting Sj-S by direct Dot-ELISA and indirect Dot-ELISA. However, the positive reaction rate was from 20% to 50% in detecting Hp-S, P-S, C-S and T-S by direct Dot-ELISA, and it was 0 in detecting these sera by indirect Dot-ELISA. Conclusions There is a high affinity between ASSP and the antibodies of Schistosoma japonicum, and the ASSP is a potential diagnoslic antigen for detection of the antibodies of Schistosoma japonicum in schistosome infected people＇s serum.