叶雪梅, 胡佳佳, 胡静. 检测食品沙门氏菌的生物传感器持久性研究[J]. 农业工程学报, 2014, 30(20): 334-338. DOI: doi:10.3969/j.issn.1002-6819.2014.20.040
    引用本文: 叶雪梅, 胡佳佳, 胡静. 检测食品沙门氏菌的生物传感器持久性研究[J]. 农业工程学报, 2014, 30(20): 334-338. DOI: doi:10.3969/j.issn.1002-6819.2014.20.040
    Ye Xuemei, Hu Jiajia, Hu Jing. Longevity analysis of biosensor for Salmonella typhimurium detection in food[J]. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE), 2014, 30(20): 334-338. DOI: doi:10.3969/j.issn.1002-6819.2014.20.040
    Citation: Ye Xuemei, Hu Jiajia, Hu Jing. Longevity analysis of biosensor for Salmonella typhimurium detection in food[J]. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE), 2014, 30(20): 334-338. DOI: doi:10.3969/j.issn.1002-6819.2014.20.040

    检测食品沙门氏菌的生物传感器持久性研究

    Longevity analysis of biosensor for Salmonella typhimurium detection in food

    • 摘要: 为了研究温度对生物传感器检测的持久性的影响,制备了用于检测食品中沙门氏菌的磁致伸缩生物传感器,以磁致伸缩膜片作为物理传感器,多克隆抗体作为生物识别元件,采用Langmuir-Blodgett(LB)技术将多克隆抗体固定在磁致伸缩膜片表面。当食品中沙门氏菌吸附在生物传感器上时,将引起其共振频率漂移。通过测试与分析磁致伸缩生物传感器共振频率漂移值,并利用扫描电子显微镜(scanning electron microscope)观察吸附了沙门氏菌的生物传感器表面,对生物传感器在25(室温)、45及65℃的持久性进行研究。结果表明:多克隆抗体磁致伸缩生物传感器与沙门氏菌的结合能力随着时间的延长逐渐降低;且温度越高,传感器的持久性越差;在25、45及65℃时,多克隆抗体磁致伸缩生物传感器的持久期分别为30、8和5 d,由此获得了多克隆抗体生物传感器在常用温度下的持久性。并结合阿伦尼乌斯方程,计算得到该生物传感器的激活能为13.024 kJ/mol。进一步证实了磁致伸缩生物传感器可用于定量检测实际溶液中沙门氏菌的浓度,表明生物传感器可应用于食品中细菌的实时快速定量检测。

       

      Abstract: Abstract: A biosensor essentially consists of two main components, a physical transducer and a biorecognition element. In this study, antibody-based magnetoelastic (ME) biosensor specific to Salmonella typhimurium has been developed by immobilizing biorecognition element onto the physical transducer. Rabbit polyclonal antibody, which was an immune system-related protein called immunoglobulins, was used as the biorecognition. A magnetostrictive platform was served as the transducer, since it offered wireless or remote detecting, which was a unique advantage over conventional sensor platforms. Amorphous magnetostrictive alloy was used as the sensor platform, which was mechanically polished using fine grit paper to reduce the thickness to 15 μm to decrease the initial mass, and magnetostrictive strips were made with the size of 5 mm×1 mm using an auto controlled, microdicing saw. Then the diced sensors were ultrasonically cleaned in methanol solution to remove grease and debris left by the dicing process. And antibody was immobilized on the magnetostrictive platform using Langmuir-Blodgett (LB) technique, in which seven monolayers containing antibody were transferred onto the magnetostrictive sensor surface using a LB film balance KSV 2200 LB. After antibody immobilization, the biosensors were divided into 3 sets and maintained in 3 constant temperature humidity chambers with temperature of 25, 45 and 65℃, respectively. Before testing, the biosensors were allowed to attain room temperature and then immersed in S. typhimurium solution with the concentration of 1×109 cfu/mL for 30 minutes to bind bacterial cells. Due to the magnetoelastic nature of the amorphous magnetostrictive alloy, the sensor exhibited a physical resonance when it underwent a time-varying magnetic field, and a shift in resonance frequency of the magnetostrictive sensor depended only on the mass change when testing environmental parameters were invariable. In this study, the environment was kept constant, and the changes in the resonance frequency of biosensors attributting to the binding of Salmonella typhimurium were recorded by HP network analyzer 8751A over the testing period, and the binding was also confirmed by scanning electron microscopy (SEM) micrographs. According to the shift in resonance frequency due to the binding of Salmonella typhimurium, the binding density bound on the biosensor was calculated. In order to confirm the binding, different regions of each sensor surface were examined and photographed using SEM, the number of cells bound to the sensor surface was directly counted from SEM images and statistically converted to an area density of bacteria attached to the sensor surface, and then compared with that calculated from the shift in resonance frequency. The longevity of the sensors at different temperatures was investigated according to the binding density of Salmonella typhimurium. Meanwhile, the activation energy was calculated using Arrhenius Equation. The results showed that at each temperature, the binding ability of antibody to S. typhimurium decreased gradually over the testing period, and the higher the temperature, the lower the longevity of the biosensor. The longevity of polyclonal antibody-based magnetostrictive sensor was about 30, 8 and 5 days at 25℃ (room temperature), 45 and 65℃, respectively. And its activation energy calculated by Arrhenius Equation was about 13.024 kJ/mol.

       

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