文章摘要
柴晓玲;郝雅茹;王佳蕊;李书国;.基于PAMAM树枝状大分子纳米免疫传感器快速检验婴幼儿奶粉中的阪崎肠杆菌[J].中国食品学报,2020,20(8):264-269
基于PAMAM树枝状大分子纳米免疫传感器快速检验婴幼儿奶粉中的阪崎肠杆菌
Rapid Determination of Enterobacter sakazakii in Infant Formula Powders by Nano-immunosensor Based on Dendrimer-PAMAM
  
DOI:
中文关键词: 婴幼儿奶粉  阪崎肠杆菌  荧光PCR  免疫传感器
英文关键词: infant formula powder  Enterobacter sakazakii  fluorescent PCR  immune sensor
基金项目:国家自然科学基金项目(20876165)
作者单位
柴晓玲;郝雅茹;王佳蕊;李书国; 河北科技大学生物科学与工程学院
 
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中文摘要:
      以聚酰胺胺树状大分子材料结合还原性石墨烯纳米复合物为电活性修饰膜层,固定于玻碳电极表面,采用物理吸附法固定化辣根过氧化物酶标记阪崎肠杆菌抗体制备成纳米免疫传感器,建立一种快速检测婴幼儿配方奶粉中阪崎肠杆菌的方法。优化的试验条件:以K3[Fe(CN)_6]为探针,过氧化氢浓度为0.5 mmol/L,pH 7.0的PBS溶液为检测底液,孵育温度37℃、孵育时间20 min。采用微分脉冲伏安法研究免疫传感器响应电流与阪崎肠杆菌浓度之间的关系。结果表明,免疫响应电流与阪崎肠杆菌浓度之间呈线性关系(ΔIpc(μA)=-1.1817 lg C+1.723,R~2=0.9989),其检出限为5.8×10~1 CFU/mL(S/N=3)。该法用于检测婴幼儿配方奶粉中的阪崎肠杆菌,其结果与国标方法(实时荧光PCR法)的数据一致,而且具有样品预处理简单、灵敏度高、快速、费用低等优点,可用于婴幼儿食品中阪崎肠杆菌的快速检测。
英文摘要:
      The immunosensor for rapid determination of Enterobacter sakazakii in infant formula powders was constructed by employing polyamidoamine(PAMAM) and reductive grapheme as electroactive nano composites to immobilize horseradish peroxidase(HRP) labelled E. sakazakii antibody on the surface of glassy carbon electrode (GCE). The optimum analytical parameters for determination of E. sakazakii as follows: using K3[Fe(CN)6] as probe, hydrogen peroxide concentration of base solution was 0.5 mmol/L, the pH of PBS solution was 7.0, incubation temperature was 37 ℃ and incubation time was 20 min. The relationship between the immune response currents and the concentration of Enterobacter sakazakii was studied by differential pulse voltammetry. The results showed that a linear relationship was found between the immune response current and the concentration of E. sakazakii (ΔIpc(μA) = 1.1817lgC-1.723, R2 = 0.9989), and the limit of detection was 5.8 × 101 CFU/mL (S/N = 3). The developed method was applied to determine E. sakazakii concentration of infant formula powders, the results were well agreed with those obtained by national standard method, and the method had the advantages of simple sample pretreatment, high sensitivity, rapid detection and low cost. Therefore, it was applicable for the rapid detection of Enterobacter sakazakii in infant formula powders.
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