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利用人免疫球蛋白制备富集金黄色葡萄球菌的免疫磁珠及优化条件;利用化学键法制备免疫磁珠,并用矩阵法和单因素法优化免疫磁珠制备条件,通过磁珠对抗体饱和吸附量及加入量相关性评价磁珠与抗体固定化程度,根据免疫磁珠的特异性、灵敏性及饱和捕捉量来考察IMBS.aureus对金葡菌的富集能力;结果显示采用MB-M型磁珠,加入质量浓度均为5 mg/L的活化剂EDC和NHS,于24℃偶联人IgG,24 h得到IMBS.aureus,其饱和吸附量为393.27μg,磁珠与人IgG加入量具有相关性,IMBS.aureus特异性好、灵敏度高(26 CFU/10 mL),0.2 mL IMBS.aureus饱和捕捉菌液浓度为106CFU/mL;通过优化,磁珠装载人IgG分子的能力提高了5倍,检测限提高了4倍,富集金葡菌的效率大大增强,成本降低,为人免疫球蛋白磁珠富集和分离金黄色葡萄球菌实际应用创造了条件。
The immunomagnetic beads enriched in Staphylococcus aureus were prepared by using human immunoglobulin and the optimization conditions were set up. The immunomagnetic beads were prepared by chemical bond method. The conditions of immunomagnetic bead preparation were optimized by matrix method and single factor method. And the amount of added to evaluate the degree of immobilization of magnetic beads and antibodies, according to the specificity of the immunomagnetic beads, sensitivity and saturation capture capacity to investigate IMBS.aureus enrichment of Staphylococcus aureus; results show that using MB-M-type magnetic Beads were added EDC and NHS with mass concentration of 5 mg / L, and IgG was coupled at 24 ℃ for 24 h to obtain IMBS.aureus with a saturated adsorption capacity of 393.27 μg. Magnetic beads were correlated with the addition of human IgG Sex, IMBS.aureus specificity, high sensitivity (26 CFU / 10 mL), 0.2 mL IMBS.aureus saturated capture bacteria concentration of 106CFU / mL; by optimization, the ability of magnetic beads to load human IgG molecules increased by 5 times, The detection limit increased by 4 times, the enrichment of S. aureus greatly enhanced the efficiency, lower costs, as human immunoglobulin beads enrichment and separation of Staphylococcus aureus practical application of the conditions.