title: Sample introduction interface for on-chip nucleic acid-based analysis of Helicobacter pylori from stool samples.
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.
authors.affiliation: Faculty of Science and Engineering, Manchester Metropolitan University, Chester Street, Manchester, M1 5GD, UK. k.shaw@mmu.ac.uk.
authors.forename: O
authors.initials: O
authors.lastname: Mosley
authors.affiliation: Faculty of Science and Engineering, Manchester Metropolitan University, Chester Street, Manchester, M1 5GD, UK. k.shaw@mmu.ac.uk.
authors.forename: L
authors.initials: L
authors.lastname: Melling
authors.affiliation: Department of Chemistry, University of Hull, Cottingham Road, Hull, HU6 7RX, UK. n.pamme@hull.ac.uk.
authors.forename: M D
authors.initials: MD
authors.lastname: Tarn
authors.affiliation: Department of Chemistry, University of Hull, Cottingham Road, Hull, HU6 7RX, UK. n.pamme@hull.ac.uk.
authors.forename: C
authors.initials: C
authors.lastname: Kemp
authors.affiliation: School of Engineering, University of Hull, Cottingham Road, Hull, HU6 7RX, UK.
authors.forename: M M N
authors.initials: MM
authors.lastname: Esfahani
authors.affiliation: Department of Chemistry, University of Hull, Cottingham Road, Hull, HU6 7RX, UK. n.pamme@hull.ac.uk.
authors.forename: N
authors.initials: N
authors.lastname: Pamme
authors.affiliation: Faculty of Science and Engineering, Manchester Metropolitan University, Chester Street, Manchester, M1 5GD, UK. k.shaw@mmu.ac.uk.
authors.forename: K J
authors.initials: KJ
authors.lastname: Shaw
mesh_terms: D004269:DNA, Bacterial / Q000032:analysis*; D005243:Feces / Q000382:microbiology*; D016480:Helicobacter pylori / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D056656:Lab-On-A-Chip Devices
chemical_list: D004269:DNA, Bacterial