Immunomagnetic nanobeads based on a streptavidin-biotin system for the highly efficient and specific separation of Listeria monocytogenes
Immunomagnetic nanobeads based on a streptavidin-biotin system for the highly efficient and specific separation of Listeria monocytogenes
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基于链霉亲和素-生物素系统的免疫磁性纳米珠，用于高效、特异性分离单核细胞增生李斯特氏菌
DOI：
10.1016/j.foodcont.2014.04.036
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发表时间：
2014-11-01
期刊：
影响因子：
6
通讯作者：
Liu, Daofeng
中科院分区：
文献类型：
--
作者：
Shan, Shan;Zhong, Ziqing;Liu, Daofeng
采用免疫磁性分离技术分离单核细胞增生李斯特菌，以缩短检测时间。优化了偶联条件和免疫磁珠（IMB）捕获工艺。结果表明，0.1 mg 180 nm免疫磁珠（IMB-S-180）与单克隆抗体偶联，链霉亲和素-生物素系统的孵育时间和分离时间分别为45 min和3 min。磁选机的分选介质和磁场强度对磁选效果有影响。当磁场强度为1.5T时，使用1 ml PBS作为分离介质，捕获效率（CE）为94.12%。IMB与LM菌株表现出高度特异性结合（CE > 90%），但与非靶细菌的结合较低（CE < 5%）。IMBs（0.1mg/m1）对绞细牛肉和牛奶样品中的LM的CE分别为25.1%和91.2%。这些结果可以作为一个指导适当选择的IMB开发的病原体分离过程。(C)2014爱思唯尔有限公司版权所有。
Immunomagnetic separation was used to decrease the assay time for Listeria monocytogenes (LM) isolation. The coupling conditions and immunomagnetic bead (IMB) capture process were optimized. Results showed that the incubation time and separation time for 0.1 mg of 180 nm immunomagnetic nanobeads (IMB-S-180) coupled with monoclonal antibody using a streptavidin-biotin system was 45 and 3 min, respectively. Magnetic separation was affected by the separation medium and magnetic density of the magnetic separator. Capture efficiency (CE) was 94.12% when the magnetic density was 1.5 T, using 1 ml of PBS as separation medium. The IMBs exhibited highly specific binding with LM strains (CE > 90%) but low binding with non-target bacteria (CE < 5%). The CE of IMBs (0.1 mg/m1) against LM in ground beef and milk samples were 25.1% and 91.2%, respectively. These results can serve as a guide for the appropriate selection of IMBs for developing isolation processes for pathogens. (C) 2014 Elsevier Ltd. All rights reserved.