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目的通过对裸磁珠、单一抗体和混合抗体包被的免疫磁珠的吸附性能的比较,建立一种快速、同时富集多种致病菌的方法。方法通过沉淀法制备Fe3O4磁珠,对其进行表面修饰,用抗体进行偶联制备成免疫磁珠,比较不同磁珠对致病菌的吸附性能。结果通过化学合成和修饰的方法制备出裸磁珠和Fe3O4/SiO2的免疫磁珠。免疫磁珠对细菌的吸附率与裸磁珠比较差异显著(t=7.55,P<0.05),平均吸附率分别为96.62%和80.89%。通过洗涤优化,免疫磁珠对目标菌的吸附率从97.03%降到93.67%,对非目标菌的吸附率从53.33%降到16.67%;裸磁珠对目标菌和非目标菌的吸附率从81%降到13.33%;免疫磁珠对单一菌和混合菌的吸附性能无显著差异(P=0.427)。结论单一抗体和混合抗体包被的免疫磁珠能特异性富集目标菌,通过富集程序优化可以达到良好的吸附效果。
OBJECTIVE To establish a rapid and simultaneous method for enrichment of multiple pathogenic bacteria by comparing the adsorption performance of magnetic beads coated with naked magnetic beads, single antibody and mixed antibody. Methods Fe3O4 magnetic beads were prepared by precipitation method, and surface modified. The magnetic beads were prepared by conjugation with antibodies, and the adsorption properties of different magnetic beads on pathogenic bacteria were compared. Results Magnetic beads and Fe3O4 / SiO2 immunomagnetic beads were prepared by chemical synthesis and modification. The adsorption rate of bacteria by magnetic beads was significantly different from that of bare magnetic beads (t = 7.55, P <0.05), and the average adsorption rates were 96.62% and 80.89% respectively. By washing optimization, the adsorption rate of the target magnetic beads to the target bacteria decreased from 97.03% to 93.67%, while the adsorption rate to non-target bacteria dropped from 53.33% to 16.67%. The adsorption rate of the target magnetic beads to the target bacteria and non-target bacteria was 81% to 13.33%. There was no significant difference (P = 0.427) between the immunomagnetic beads adsorption of single bacteria and mixed bacteria. Conclusion The single antibody and the mixed antibody coated immunomagnetic beads can specifically enrich the target bacteria, and the enrichment program can optimize the adsorption effect.