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目的建立一种无需仪器、操作简单,具较好敏感性和特异性的适用于基层快速检测广州管圆线虫感染的胶体金免疫层析方法。方法将重组广州管圆线虫五期幼虫抗原Ac11蛋白为检测点膜抗原,羊抗兔IgG喷涂在质控线,与胶体金标记兔抗小鼠单抗组装成广州管圆线虫胶体金免疫层析法检测试剂条,用于检测阳性小鼠血清特异性抗体。试验设阴性小鼠血清为对照。结果确定试剂条的检测条件为:检测线重组抗原Ac11用量为4mg/ml(喷量1μl/cm),质控线羊抗兔IgG用量为1.87mg/ml(喷量1μl/cm),胶体金标记兔抗小鼠单抗采用吸光度(A)值为5的喷涂量(喷量2μl/cm)。用该方法检测小鼠血清抗广州管线虫抗体,其敏感性为100%(10/10),特异性为100%(10/10)。结论胶体金免疫层析法检测广州管圆线虫抗体具有较好的敏感性和特异性,经进一步完善后有望用于广州管圆线虫病的临床快速筛查和流行病学调查。
OBJECTIVE: To establish a colloidal gold immunochromatography method for the rapid detection of A. cercariae infection at the grass-roots level without any instrumentation, simple operation, high sensitivity and specificity. Methods Ac11 protein of Acinetobacter nocturnalis larvae was used as detection membrane antigen, goat anti-rabbit IgG was sprayed on the control line, and colloidal gold labeled rabbit anti-mouse monoclonal antibody was assembled into Agrobacterium tumefaciens France test strips for the detection of positive mouse serum-specific antibodies. Negative mouse serum test as a control. Results The conditions for the detection of the reagent strip were as follows: the amount of Ac11 in the test line was 4mg / ml (spray volume 1μl / cm); the amount of goat anti-rabbit IgG in control line was 1.87mg / ml The labeled rabbit anti-mouse mAb was sprayed at an absorbance (A) value of 5 (spray volume 2 μl / cm). Using this method to detect mouse serum anti-nematode antibodies, the sensitivity was 100% (10/10) and the specificity was 100% (10/10). Conclusion The colloidal gold immunochromatography assay has good sensitivity and specificity for detecting Agrobacterium tumefaciens. After further refinement, it is expected to be used in rapid screening and epidemiological investigation of GAV in Guangzhou.