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Lab Chip. 2016 May 24;16(11):2108-15. doi: 10.1039/c6lc00228e.

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

Author information

1
Faculty of Science and Engineering, Manchester Metropolitan University, Chester Street, Manchester, M1 5GD, UK. k.shaw@mmu.ac.uk.
2
Department of Chemistry, University of Hull, Cottingham Road, Hull, HU6 7RX, UK. n.pamme@hull.ac.uk.
3
School of Engineering, University of Hull, Cottingham Road, Hull, HU6 7RX, UK.

Abstract

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.

PMID:
27164181
DOI:
10.1039/c6lc00228e
[Indexed for MEDLINE]

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