基于NMR纳米探针的生物传感器用于快速检测沙门氏菌
Biological sensors based on nano magnetic beads applied in rapid detection of salmonella
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摘要: 通过将沙门氏菌单抗与羧基磁珠偶联制备免疫磁珠,并以此为NMR分子探针,以免疫磁珠为生物传感器,特异性地捕获并检测出样品中的致病菌,从而建立一种更快的检测沙门氏菌的方法。实验将得到的不同样品溶液放入核磁共振仪中检测自旋-自旋弛豫时间(T2)值,考察不同条件对T2值的影响。通过实验发现,免疫磁珠的使用量,缓冲溶液的选择,捕获时间的长短都会对样品T2值产生影响。通过优化实验,分别绘制出不同条件下T2值的曲线变化,并得出最佳捕获条件。在捕获缓冲溶液为磷酸盐缓冲溶液PBS(0.01 mol/L,p H7.4),免疫磁珠使用量为50μg,混匀捕获时间为1.5 h的条件下,核磁共振仪测得的样品T2值曲线最佳。其检测线性范围为104107CFU/m L。本研究为沙门氏菌的检测提供了新的方法和途径,缩短了检测时间和工序,并且为低场核磁共振技术研究应用开辟了广阔的空间。Abstract: By coupling salmonella monoclonal antibody with carboxyl magnetic beads,immunomagnetic beads were produced,which were used as NMR molecular probes and biological sensors,pathogenic bacteria specially captured and detected in the samples so as to create a more rapid method to detect salmonella.Different sample solutions were put into the nuclear magnetic resonance apparatus to detect the spin relaxation time( T2) and examine the influence of different conditions to the value of T2. The experiment found that the sample value of T2 was influenced by the volume of immunomagnetic beads,the choice of buffer solution and the time of capturing.Through optimization experiment,curve variations T2 under different conditions were drawn with a result of ideal capturing conditions.Under the conditions that the capturing buffer solution was phosphate buffer solution( PBS)( 0.01 mol/L,p H7.4),the volume of immunomagnetic beads was 50 μg and the blending capturing time was 1.5 h,the curve of the sample value of T2 was perfect,detection linear range being 10~410~7CFU / m L.A new method and approach was provided by this research,which was helpful to shorten the detection time and process and opened an immense space for the study of low field nuclear magnetic resonance.