Yang Liju, Li Yanbin
Department of Biological and Agricultural Engineering, Center of Excellence for Poultry Science, University of Arkansas, Hall 203, Fayetteville, AR 72701, USA.
J Microbiol Methods. 2006 Jan;64(1):9-16. doi: 10.1016/j.mimet.2005.04.022. Epub 2005 Jun 2.
In this study, we demonstrated the use of a general medium--brain heart infusion (BHI) broth that is not specifically formulated for impedance measurement, to achieve detectable impedance signals by using an interdigitated microelectrode (IME) with capacitance measurement at low frequencies. Anti-Salmonella antibody coated immunomagnetic beads were used to separate S. typhimurium from samples to provide the selectivity to this method. From analysis based on the equivalent circuit of the IME system, we found that the impedance change in BHI broth resulting from the growth of Salmonella was indeed the change in the double layer capacitance and could be monitored at 10 Hz using the IME. The results indicated that medium modification to improve impedance signal is not necessary with this IME system. However, effective immunological separation for the target organism is required for the selectivity when non-selective media are used. This finding provides a more flexible option of medium in impedance methods, which may provide opportunities to test those species of bacteria that have no suitable conductance growth medium. The detection time, t(d), was obtained from the impedance growth curve (impedance against bacterial growth time) at 10 Hz at the point where the impedance started to change. A linear relationship between the detection time and the logarithmic value of the initial cell number (N) was found in the Salmonella cell number ranging from 10(1) to 10(6) cfu/ml. The regression equation was t(d) = -1.22Log N + 8.90, with R2 = 0.95. The detection times for the initial cell number of 10(1) CFU/ml and 10(6) CFU/ml are 8 h and 1.5 h, respectively. This method is more sensitive than impedance methods using conventional electrodes.
在本研究中,我们展示了使用一种并非专门为阻抗测量配制的通用培养基——脑心浸液(BHI)肉汤,通过使用具有低频电容测量功能的叉指微电极(IME)来实现可检测的阻抗信号。用抗沙门氏菌抗体包被的免疫磁珠从样品中分离鼠伤寒沙门氏菌,以赋予该方法选择性。基于IME系统的等效电路分析,我们发现沙门氏菌生长导致的BHI肉汤中的阻抗变化实际上是双层电容的变化,并且可以使用IME在10 Hz下进行监测。结果表明,对于该IME系统,无需对培养基进行改良以改善阻抗信号。然而,当使用非选择性培养基时,为实现选择性,需要对目标生物体进行有效的免疫分离。这一发现为阻抗法提供了一种更灵活的培养基选择,可能为检测那些没有合适电导生长培养基的细菌种类提供机会。检测时间t(d)是从10 Hz下的阻抗生长曲线(阻抗相对于细菌生长时间)中阻抗开始变化的点获得的。在沙门氏菌细胞数为10(1)至10(6) cfu/ml的范围内,发现检测时间与初始细胞数(N)的对数值之间存在线性关系。回归方程为t(d) = -1.22Log N + 8.90,R2 = 0.95。初始细胞数为10(1) CFU/ml和10(6) CFU/ml时的检测时间分别为8小时和1.5小时。该方法比使用传统电极的阻抗法更灵敏。