Sample introduction interface for on-chip nucleic acid-based analysis of: Helicobacter pylori from stool samples

Mosley, O, Melling, L ORCID: https://orcid.org/0000-0001-7690-5429, Tarn, MD, Kemp, C, Esfahani, MMN, Pamme, N and Shaw, KJ (2016) Sample introduction interface for on-chip nucleic acid-based analysis of: Helicobacter pylori from stool samples. Lab on a Chip: miniaturisation for chemistry, physics, biology, materials science and bioengineering, 16 (11). pp. 2108-2115. ISSN 1473-0197

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Abstract

© The Royal Society of Chemistry 2016. Despite recent advances in microfluidic-based integrated diagnostic systems, the sample introduction interface, especially with regards to large volume samples, has often been neglected. We present a sample introduction interface that allows direct on-chip processing of crude stool samples for the detection of Helicobacter pylori (H. pylori). The principle of IFAST (immiscible filtration assisted by surface tension) was adapted to include a large volume sample chamber with a septum-based interface for stool sample introduction. Solid chaotropic salt and dry superparamagnetic particles (PMPs) could be stored on-chip and reconstituted upon sample addition, simplifying the process of release of DNA from H. pylori cells and its binding to the PMPs. Finally, the PMPs were pulled via a magnet through a washing chamber containing an immiscible oil solution and into an elution chamber where the DNA was released into aqueous media for subsequent analysis. The entire process required only 7 min while enabling a 40-fold reduction in working volume from crude biological samples. The combination of a real-world interface and rapid DNA extraction offers the potential for the methodology to be used in point-of-care (POC) devices.