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

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。

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

BACKGROUND: 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 IgM, 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 1 ug/ml, and the suitable stable concentration (SSC) was 1.5 ug/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.