Grossman H L, Myers W R, Vreeland V J, Bruehl R, Alper M D, Bertozzi C R, Clarke John
Department of Physics, Howard Hughes Medical Institute, University of California, Berkeley, CA 94720, USA.
Proc Natl Acad Sci U S A. 2004 Jan 6;101(1):129-34. doi: 10.1073/pnas.0307128101. Epub 2003 Dec 19.
We demonstrate a technique for detecting magnetically labeled Listeria monocytogenes and for measuring the binding rate between antibody-linked magnetic particles and bacteria. This sensitive assay quantifies specific bacteria in a sample without the need to immobilize them or wash away unbound magnetic particles. In the measurement, we add 50-nm-diameter superparamagnetic magnetite particles, coated with antibodies, to an aqueous sample containing L. monocytogenes. We apply a pulsed magnetic field to align the magnetic dipole moments and use a high-transition temperature superconducting quantum interference device, an extremely sensitive detector of magnetic flux, to measure the magnetic relaxation signal when the field is turned off. Unbound particles randomize direction by Brownian rotation too quickly to be detected. In contrast, particles bound to L. monocytogenes are effectively immobilized and relax in about 1 s by rotation of the internal dipole moment. This Néel relaxation process is detected by the superconducting quantum interference device. The measurements indicate a detection limit of (5.6 +/- 1.1) x 10(6) L. monocytogenes in our sample volume of 20 microl. If the sample volume were reduced to 1 nl, we estimate that the detection limit could be improved to 230 +/- 40 L. monocytogenes cells. Time-resolved measurements yield the binding rate between the particles and bacteria.
我们展示了一种检测磁性标记的单核细胞增生李斯特菌以及测量抗体连接的磁性颗粒与细菌之间结合速率的技术。这种灵敏的检测方法可对样品中的特定细菌进行定量,无需固定细菌或洗去未结合的磁性颗粒。在测量过程中,我们将涂有抗体的直径50纳米的超顺磁性磁铁矿颗粒添加到含有单核细胞增生李斯特菌的水性样品中。我们施加脉冲磁场使磁偶极矩排列整齐,并使用高转变温度超导量子干涉装置(一种极其灵敏的磁通量探测器)在磁场关闭时测量磁弛豫信号。未结合的颗粒通过布朗旋转随机化方向太快而无法被检测到。相比之下,与单核细胞增生李斯特菌结合的颗粒被有效固定,并通过内部偶极矩的旋转在大约1秒内弛豫。这种尼尔弛豫过程由超导量子干涉装置检测到。测量结果表明,在我们20微升的样品体积中,检测限为(5.6 +/- 1.1) x 10(6)个单核细胞增生李斯特菌。如果样品体积减少到1纳升,我们估计检测限可提高到230 +/- 40个单核细胞增生李斯特菌细胞。时间分辨测量得出颗粒与细菌之间的结合速率。