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目的研制一种同时适于基层医院与兽医部门使用的简便快速检测囊虫特异抗体的新方法。方法以SPA标记胶体金并固定在玻璃纤维纸上,囊虫特异抗原与兔IgG固化在NC膜上,制备可以检测囊虫特异抗体的SPA胶体金试纸条。用该试纸条检测人或猪囊虫病阳性、阴性血清以及相关疾病血清,验证方法的敏感性、特异性、重复性、可操作性以及试剂的稳定性。结果每毫升胶体金的标记SPA的最适量为5.5μg,囊虫特异抗原与兔IgG标记NC膜的最佳浓度分别为1.22mg/ml与0.97mg/ml。用SPA胶体金试纸条检测非疫区健康猪血清、人囊虫病阴性参考血清、包虫病人血清、弓形虫病人血清以及旋毛虫病人血清,结果均为阴性;检测囊虫病人血清与病猪血清,均为阳性;与I-ELISA比较,检测结果符合率100%。检测过程为2~10 min,该纸条在25℃保存2个月后的检测效果无明显变化。结论研制的SPA胶体金试纸条检测囊虫特异抗体,简便、快速、敏感、特异,试剂稳定,适于各级医院与兽医部门进行囊虫病的诊断和流行病学调查。
Objective To develop a new method for rapid and simple detection of specific antibodies against cysticercosis for both primary hospitals and veterinary departments. METHODS: Colloidal gold was labeled with SPA and immobilized on glass fiber paper. Cyclosporidium specific antigen and rabbit IgG were immobilized on NC membrane to prepare SPA colloidal gold test strips for detection of cysticercus-specific antibodies. This test strip is used to test the sensitivity, specificity, repeatability, operability and reagent stability of human or swine cystosis-positive, negative serum and related diseases serums. Results The optimal amount of labeled SPA per ml of colloidal gold was 5.5 μg. The optimum concentration of cystathion specific antigen and rabbit IgG-labeled NC membrane were 1.22 mg / ml and 0.97 mg / ml, respectively. The results showed that all the samples were negative with the serum of colloidal gold test strips, the serum of the healthy pigs in non-infected area, the negative reference serum of human cysticercosis, the serum of hydatid disease patients and the serum of patients with Toxoplasma gondii, Serum and pig serum, were positive; compared with I-ELISA, the test results in line with the rate of 100%. The detection process is 2 ~ 10 min, the paper did not change significantly after 2 months of storage at 25 ℃. Conclusion The gel colloidal gold test strip developed for the detection of cysticercosis specific antibodies is simple, rapid, sensitive, specific and stable. It is suitable for the diagnosis and epidemiological investigation of cysticercosis in hospitals and veterinary departments at all levels.