Micro corona discharge based cell lysis method suitable for inhibitor resistant bacterial sensing systems

Abstract

We demonstrated a bacterial cell lysis method suitable for inhibitor resistant bacterial detection systems where purification of extracted DNA is not necessary. The presented method potentially improves the field portability of such systems. It allows cell lysis and DNA extraction to be performed without the use of bead mill, sonication, thermal cycling, additional reagents or enzymes. Bacterial cell lysis is achieved in a single step by pumping ozone generated by a micro corona discharge into the bacterial sample. The results with Pseudomonas putida as the target bacteria showed that it was capable of achieving 98.5 +/- 0.2% lysis (normalized to 1 min of sonication at 10 W) after 10 min of treatment at a flow rate of 38 ml/min and an applied voltage of 2000 V. By increasing the treatment duration, flow rate and applied voltage, the normalized % lysis could be increased. In addition, continuous and pulsed treatments yield similar normalized % cell lysis. (C) 2015 Elsevier B.V. All rights reserved.