滴金免疫测定法用于检测囊虫病患者循环抗原的研究

目的　建立一种敏感、快速、简便的囊虫病诊断方法。方法　应用两株抗猪囊尾蚴囊液抗原的单克隆抗体(McAb) ,一株滴于硝酸纤维素膜上 ,另一株与胶体金结合为探针 ,建立了检测囊虫病患者血清中循环抗原 (CA)的滴金免疫测定法 ,抗原、抗体通过渗滤在膜上进行反应 ,10min内即可用肉眼观察结果。对不同稀释度的猪囊尾蚴囊液抗原进行检测 ,确定本法的最低抗原检出量 (0 .4 5ng/ml)。将本法与ELISA作了比较。 结果　对 78例囊虫病患者血清进行检测 ,循环抗原阳性率为 87.18%。其中活动型脑囊虫病患者 5 2例 ,循环抗原阳性率为 98.0 8% ;非活动型脑囊虫病患者 2 0例 ,循环抗原阳性率为 6 5 .0 0 % ;单纯皮肌型囊虫病患者 6例 ,循环抗原阳性率为 6 6 .6 7%。 4 0份健康人血清仅有 1例 (2 .5 0 % )出现假阳性 ,与10例华支睾吸虫病患者血清、15例血吸虫病患者血清及 2 3例非囊虫病的脑部疾病患者血清均无交叉反应。该方法检出最低猪囊尾蚴囊液抗原量为 0 .4 5ng/ml。本法与ELISA的符合率为 98.19%。结论　DIGFA简便、快速 ,结果客观 ,对囊虫病特别是活动型脑囊虫病敏感性高 ,特异性强 ,值得进一步推广应用。

The use of DIGFA for the detection of circulating antigens in sera of patients with cysticercosis

Aim To set up a method that can diagnose cysticercosis rapidly and sensitively.Methods Two McAbs against cyst fluid antigen of cysticercus cellulosae,one absorbed on nitrocellulose membrane,another conjugated to colloidalgold,were used to develop a dot immunogold filtration assay(DIGFA)for the detection of circulating antigens(CA) in sera of patients with cysticercosis.Antigen-antibody reaction was completed on the membrane during the filtration process.The result could be read with the naked eye within ten minutes.DIGFA and ELISA were compared. Results 68 of 78 sera from cysticercosis were positive.The positive rat was 87.18%.Of the 78 sera from cysticercosis,51 of 52(98.08%)sera from active neurocysticercosis,13 of 20(65.00%)sera from inactive neurocysticercosis and 4 of 6( 66.67%)sera from cysticercosis of simple subcutaneous nodules type were positive.Only one out of 40 sera from normal population showed positive.None of 10 sera from patients with clonorchiasis,15 sera from patients with schistosomiasis and 23 sera from patients with other diseases of the central nervous system was positive.The minimal detectable level of DIGFA was 0.45ng/ml.DIGFA and ELISA had a similar sensitivity and specificity.The coincidence rat was 98.19%.However,DIGFA was simpler and easier to conduct and required no instrument.Conclusion DIGFA is suitable for diagnosis of cysticercosis,especially for active neurocysticercosis.