Evaluation of a microfluidic chip system for preparation of bacterial DNA from swabs, air, and surface water samples.

The detection of bacterial pathogens from complex sample matrices by PCR requires efficient DNA extraction. In this study, a protocol for extraction and purification of DNA from swabs, air, and water samples using a microfluidic chip system was established. The optimized protocol includes a combination of thermal, chemical and enzymatic lysis followed by chip-based DNA purification using magnetic particles. The procedure was tested using Gram-positive Bacillus thuringiensis Berliner var. kurstaki as a model organism for Bacillus anthracis and the attenuated live vaccine strain of Francisella tularensis subsp. holarctica as Gram-negative bacterium. The detection limits corresponded to 103 genome equivalents per milliliter (GE/ml) for surface water samples spiked with F. tularensis and 102 GE/ml for B. thuringiensis. In air, 10 GE of F. tularensis per 10 L and 1 GE of B. thuringiensis per 10 L were detectable. For swab samples obtained from artificially contaminated surfaces the detection limits were 4 × 103 GE/cm2 for F. tularensis and 4 × 102 GE/cm2 for B. thuringiensis. Suitability of the chip-assisted procedure for DNA preparation of real samples was demonstrated using livestock samples. The presence of thermophilic Campylobacter spp. DNA could be confirmed in air samples collected on pig and broiler farms.

Building up longitudinal concentration gradients in shallow microchannels.

We demonstrate a compact and low consumption (60 nL) method for generating concentration gradients along microchannels with shallow parabolic cross-sections. The regimes of dispersion at work in such systems and the resulting concentration fields are described theoretically and experimentally. Experiments are performed in PDMS (polydimethylsiloxane) microchannels actuated by integrated valves. Detailed comparison between theory and experiment for the "short time" and "long time" regimes leads to excellent agreement. The system is used to successfully set up a series of isolated microchambers with mixtures of increasing solute concentrations, which may be a first step towards devices for screening.