Journal of Pure and Applied MicrobiologyVol. 11 No. 3

Multi-frequency Microchannel Electrical Impedance(m-EIS) Method for the Rapid Detection of Proliferating Microorganisms, and their Rapid Quantification

Sachidevi Puttaswamy1, Byung-Doo Lee1, Ashley Jurgensmeyer1, Anne Baumstummler2, Kathleen Souza3 and Shramik Sengupta1

1Department of Bioengineering, University of Missouri, 1406, E. Rollins Road, 165 AEB, Columbia MO 65211-5200. 2Department of Predevelopment - Technology - Collaboration, BioMonitoring, Lab Solutions, MilliporeSigma 39 Route Industrielle de la Hardt; 67120 Molsheim, France. 3Department of Virology and Microbiological Sciences, MilliporeSigma 80 Ashby Rd, Bedford, MA 01886.

Received on 13 June 2017 and accepted on 24 August 2017

 

ABSTRACT

Existing culture-based instruments for detecting/quantifying proliferating bacteria in suspensions (BACTECTM, BacT/AlertTM, RABITTM etc.) do so based on changes observed in the physical/chemical properties of media (O2/CO2 levels, pH etc.) due to bacterial metabolism. Given the limited metabolic-rate of individual bacterium, they have a “threshold-concentration” of ~107-108CFU/ml, and Times to Detection (TTDs) of 12 hours or longer for low initial loads (<100CFU/ml). We recently developed a method that tracks microbial proliferation in suspensions by monitoring the degree of cell polarization of live microorganisms. In the presence of an AC electric field, there occurs a build-up of charge at the microbial membrane, causing them to act like capacitors. As microorganisms multiply, there occurs a corresponding increase in charges stored in the suspension (“bulk-capacitance”), and this increase in bulk-capacitance serves as our “signature” for presence of live microorganisms. In this study, we explain the theory underlying our approach, establish its applicability to a variety of microorganisms, showing that the “Threshold-Concentration” (nT) for detection is ~103-104CFU/ml, and TTDs are a function of the initial-load(n0) and doubling-time(tD) of the microorganism TTD=1.443*tD*ln(nT/n0) and show that the method can be adapted to obtain the “Most Probable Number” (MPN) of coliforms within 6hrs (vs. >24hrs for existing methods).

Keywords : Viable bacteria; MPN; Rapid Detection; Automated Culture Systems; Bacteria detection; Microfluidics.