School of Medicine, Istanbul Kemerburgaz University, 34217, Istanbul, Turkey.
Appl Microbiol Biotechnol. 2013 Nov;97(21):9541-51. doi: 10.1007/s00253-013-5231-0. Epub 2013 Sep 19.
We report an immuno-magnetic separation system developed by the immobilization of pathogen-specific antibodies on the core-shell magnetic beads. The magnetic beads were grafted with glycidylmethacrylate (GMA) using surface-initiated atom transfer radical polymerization (SI-ATRP). For immuno-magnetic separation (IMS) of target bacterial cells from others, antibodies for Escherichia coli and Salmonella enterica serovar Typhimurium cells were immobilized on the magnetic beads via glutaraldehyde coupling reaction. Our IMS system successfully separated Salmonella cells when the concentrations of target (i.e., Salmonella) and interfering (i.e., E. coli) cells were at the same level. Polymerase chain reaction (PCR) assays amplifying the rfb/rfbE region of the E. coli genome and a 647-bp fragment of the invA region of Salmonella were performed as the specific selection to accurately confirm the presence of E. coli and Salmonella, respectively. IMS and multiplex PCR methods can be used for specific and quantitative detection of pathogens from food samples. Thus, this study developed a reliable and direct system for rapid detection of Salmonella and E. coli in food samples. In addition, IMS method could be easily adapted to detect other pathogens by selecting the pertinent antibody.
我们报告了一种免疫磁分离系统，该系统通过将病原体特异性抗体固定在核壳型磁性珠上而开发。使用表面引发原子转移自由基聚合（SI-ATRP）将磁性珠接枝上甲基丙烯酸缩水甘油酯（GMA）。为了从其他细菌细胞中免疫磁分离（IMS）目标细菌细胞，将针对大肠杆菌和鼠伤寒沙门氏菌血清型的抗体通过戊二醛偶联反应固定在磁性珠上。当目标（即沙门氏菌）和干扰（即大肠杆菌）细胞的浓度相同时，我们的 IMS 系统成功地分离了沙门氏菌细胞。聚合酶链反应（PCR）扩增大肠杆菌基因组的 rfb/rfbE 区域和沙门氏菌 invA 区域的 647-bp 片段作为特定选择，分别准确确认大肠杆菌和沙门氏菌的存在。IMS 和多重 PCR 方法可用于从食品样品中特异性和定量检测病原体。因此，本研究开发了一种可靠且直接的系统，用于快速检测食品样品中的沙门氏菌和大肠杆菌。此外，通过选择相关抗体，IMS 方法可以很容易地适应检测其他病原体。
