应用胶体金免疫层析法快速检测恶性疟原虫中HRPⅡ抗原研究

A RAPID IMMUNOCHROMATOGRAPHIC TEST (ICT) FOR DIAGNOSIS OF PLASMODIUM FALCIPARUM

目的建立一种简易快速、适用于基层的胶体金免疫层析法 (ICT)用于检测恶性疟原虫中HRPⅡ抗原 ,判定是否现症感染。方法采用柠檬酸三钠还原法制备胶体金颗粒、标记HRPⅡ单克隆抗体 ,制成免疫测试条。血中HRPⅡ抗原与测试条上金标记单克隆抗体结合后沿着硝酸醋酸纤维素膜移动 ,与膜上的固相特异性单克隆抗体结合形成肉眼可见的褐红色线。结果 ICT测试条只与HRPⅡ抗原有特异性反应 ,与间日疟无交叉反应。与显微镜检比较特异性为 10 0 %。用该法检测体外培养的恶性疟原虫HRPⅡ抗原 ,其灵敏度在体外培养实验中 ,当红细胞感染虫体为 1%时使用 5 μl血稀释 10 0倍仍能检测出来。该法检测 30份用病原学确诊的恶性疟原虫和 2 0份间日疟标本 ,检出率为 10 0 % ,与间日疟无交叉反应。结论 ICT检测血中恶性疟原虫中HRPⅡ抗原特异性强、灵敏度高、简便快速 ,无需特殊仪器设备 。

Aim To evaluate the rapid dipstick test in the diagnosis of P.falciparum infection in human.Methods Assess the sensitivity and specificity of the rapid immunodiagnostic test based on detection of P.falciparum histidine rich protein-Ⅱ (PFHRP-Ⅱ) in blood,which was antigen-capture assay based on monoclonal atibodies of the P.falciparum histidine-rich protein Ⅱ(HRP Ⅱ).Results Evaluation in 50 patients sample showed that the ICT had 100% sensitivity when using microscopy based on gold standard.There was no corss-reactivity with P.vivax.Conclusions The test is a valuable diagnostic tool for falciparum malaria,especially in emergency/field situations requiring rapid diagnosis.