Duan Hui-Li, Shen Zhi-Qiang, Wang Xin-Wei, Chao Fu-Huan, Li Jun-Wen
Department of Environmental Health, Institute of Hygiene and Environmental Medicine, Tianjin 300050, China.
World J Gastroenterol. 2005 Jun 28;11(24):3660-4. doi: 10.3748/wjg.v11.i24.3660.
To prepare a kind of magnetic iron-dextran nanoparticles that was coated with anti-E.coli O157:H7 IgG, analyze its application conditions, and try to use it to isolate E.coli O157:H7 from foods.
Magnetic iron-dextran nanoparticles were prepared by the reaction of a mixture of ferric and ferrous ions with dextran polymers under alkaline conditions. The particles were coated with antiserum against E.coli O157:H7 by the periodate oxidation-borohydride reduction procedure. The oxidation time, amount of antibody coating the particles, amount of nanoparticles, incubation time and isolation time were varied to determine their effects on recovery of the organisms. Finally, the optimum conditions for isolating E.coli O157:H7 from food samples were established.
E.coli O157:H7 can be isolated from samples within 15 min with the sensitivity of 10(1) CFU/mL or even less. In the presence of 10(8) CFU/mL of other organisms, the sensitivity is 10(1)-10(2) CFU/mL. Nonspecific binding of other bacteria to the particles was not observed. Two and a half hours of enrichment is enough for the particles to detect the target from the food samples inoculated with 1 CFU/g.
Isolation of target bacteria by immunomagnetic nanoparticles is an efficient method with high sensitivity and specificity. The technique is so simple that it can be operated in lab and field even by untrained personnel.
制备一种包被抗大肠杆菌O157:H7 IgG的磁性铁-葡聚糖纳米颗粒,分析其应用条件,并尝试用其从食品中分离大肠杆菌O157:H7。
在碱性条件下,通过三价铁离子和二价铁离子的混合物与葡聚糖聚合物反应制备磁性铁-葡聚糖纳米颗粒。采用高碘酸盐氧化-硼氢化钠还原法用抗大肠杆菌O157:H7抗血清包被颗粒。改变氧化时间、包被颗粒的抗体量、纳米颗粒量、孵育时间和分离时间,以确定它们对微生物回收率的影响。最后,确定了从食品样品中分离大肠杆菌O157:H7的最佳条件。
可在15分钟内从样品中分离出大肠杆菌O157:H7,灵敏度可达10(1) CFU/mL甚至更低。在存在10(8) CFU/mL其他微生物的情况下,灵敏度为10(1)-10(2) CFU/mL。未观察到其他细菌与颗粒的非特异性结合。对于接种了1 CFU/g的食品样品,颗粒进行两个半小时的富集就足以检测到目标菌。
用免疫磁性纳米颗粒分离目标细菌是一种高效、灵敏度高且特异性强的方法。该技术非常简单,即使未经培训的人员也可在实验室和现场操作。