Detection of viable Salmonella using microelectrode-based capacitance measurement coupled with immunomagnetic separation.
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.
Yang
Liju
L
Department of Biological and Agricultural Engineering, Center of Excellence for Poultry Science, University of Arkansas, Hall 203, Fayetteville, AR 72701, USA.
Li
Yanbin
Y
D015374:Biosensing Techniques / Q000379:methods*; D002138:Calibration; D003470:Culture Media / Q000378:metabolism; D025802:Electric Capacitance; D017097:Electric Impedance; D005516:Food Microbiology; D018189:Immunomagnetic Separation / Q000379:methods*; D008839:Microelectrodes; D012486:Salmonella typhimurium / Q000254:growth & development / Q000302:isolation & purification* / Q000502:physiology
D003470:Culture Media