Rapid Detection of Enterobacter Sakazakii in milk Powder using amino modified chitosan immunomagnetic beads.
Rapid Detection of Enterobacter Sakazakii in milk Powder using amino modified chitosan immunomagnetic beads.
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DOI：
10.1016/j.ijbiomac.2016.09.024
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发表时间：
2016-12
影响因子：
8.2
通讯作者：
Yinglian Zhu;Dongfeng Wang
中科院分区：
文献类型：
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作者：
Yinglian Zhu;Dongfeng Wang
以环氧氯丙烷和乙二胺为修饰剂，将天然高分子材料壳聚糖的羟基改性为氨基，再以戊二醛为交联剂与坂崎肠杆菌多克隆抗体偶联，制备壳聚糖免疫磁珠（CIBs）。采用磁性测试、FTIR、SEM和XRD等技术对偶联前的磁珠进行了表征。本试验首次将具有良好生物化学性质的天然多糖-壳聚糖用于免疫磁珠的合成。本文首次建立了将微球与显色介质相结合的快速检测方法。可大大提高检测特异性和工作效率。最大捕获量为1 × 106 cfu/g，检测灵敏度为4cfu/g。结果表明，该方法简便、特异、灵敏，可用于食品基质中坂崎肠杆菌的快速检测。总分析时间仅为25 h左右，大大缩短了检测时间。该方法不仅为免疫磁珠的制备技术提供了新的思路，也为食品安全中的免疫检测技术提供了新的思路。
Chitosan immunomagnetic beads (CIBs) were first prepared through converting hydroxyl groups of natural polymer material-chitosan into amino groups using epichlorohydrin and ethylenediamine as modification agent and then coupling with polyclonal antibodies ofEnterobacter sakazakiiusing glutaraldehyde as cross-linking agent. The beads before coupling with antibodies were characterized by magnetic property measurement, FTIR, SEM and XRD technologies. In the assay a natural polysaccharide-chitosan, which has good biological and chemical properties such as non-toxicity, biocompatibility and high chemical reactivity was first used for synthesis of immunomagnetic beads. The detection method first established in this paper that combined the beads with chromogenic medium together to rapid detectE. sakazakiiin milk powder could greatly improve the detection specificity and working efficiency. The beads exhibited a maximum capturing capacity of 1 × 106cfu/g with the detection sensitivity of 4 cfu/g. The results demonstrate that the assay is a straightforward, specific and sensitive alternative for rapid detection ofE.sakazakiiin food matrix. The total analysis time was as little as about 25 h, which greatly shorten the detection time. The method can provides new ideas not only to preparation technique of immunomagnetic beads but to imunne detection technique in food safety.