柯萨奇病毒IgM抗体胶体金免疫层析快速诊断试纸条的研制

Research on development of gold immunochromagraphic assay test kit for serum Coxsackievirus IgM antibody

目的建立一种快速检测柯萨奇病毒IgM抗体的胶体金免疫层析试纸条,并优化制备中各关键步骤的实验条件。方法以柠檬酸三钠还原法制备20nm的胶体金溶液,标记羊抗人IgM,制备免疫胶体金复合物,组装胶体金免疫层析快速诊断试纸条。结果用柠檬酸还原法制备的20nm胶体金溶液呈亮红色。胶体金标记羊抗人IgM最低稳定量是1μg/ml,最适稳定量为1.5μg/ml。最适pH为8.2。血清标本检测结果与进口ELISA试剂盒比较差异无统计学意义。结论胶体金免疫层析试纸条制备质量不但与抗原抗体的质量、层析材料的选择、胶体金的制备与标记等因素密切相关,而且缓冲系统、辅助添加剂的选择与优化也非常重要。

Objective To develop gold immunochromatographic assay （GICA） test kit for serum Coxsackievirus IgM antibody and optimize the key experiment conditions. Methods Colloidal gold particles of 20 nm were prepared and coupled with sheep anti-human lgM, the gold immunochromatography assay test kit was setup for serum Coxsackievirus IgM antibody. Results The spheroidal colloid gold particles of 20 nm showed bright red in color. The minimal stable concentration （MSC） of gold-sheep anti-human IgM was 1μg/ml,and the suitable stable concentration （SSC） was 1.5 μg/ml. The pH 8.2 was appropriate; 30 sera samples were tested by GICA and ELISA, there was no significant difference between the two methods. Conclusion The quality of gold immunochromatography assay test kit is associated with such factors as the quality and quantity of antigen or antibody, colloid gold particles, buffers, etc.