title: Towards a "Sample-In, Answer-Out" Point-of-Care Platform for Nucleic Acid Extraction and Amplification: Using an HPV E6/E7 mRNA Model System.
abstract: The paper presents the development of a "proof-of-principle" hands-free and self-contained diagnostic platform for detection of human papillomavirus (HPV) E6/E7 mRNA in clinical specimens. The automated platform performs chip-based sample preconcentration, nucleic acid extraction, amplification, and real-time fluorescent detection with minimal user interfacing. It consists of two modular prototypes, one for sample preparation and one for amplification and detection; however, a common interface is available to facilitate later integration into one single module. Nucleic acid extracts (n = 28) from cervical cytology specimens extracted on the sample preparation chip were tested using the PreTect HPV-Proofer and achieved an overall detection rate for HPV across all dilutions of 50%-85.7%. A subset of 6 clinical samples extracted on the sample preparation chip module was chosen for complete validation on the NASBA chip module. For 4 of the samples, a 100% amplification for HPV 16 or 33 was obtained at the 1 : 10 dilution for microfluidic channels that filled correctly. The modules of a "sample-in, answer-out" diagnostic platform have been demonstrated from clinical sample input through sample preparation, amplification and final detection.
authors.affiliation: NorChip AS, Industriveien 8, 3490 Klokkarstua, Norway.
authors.forename: Anja
authors.initials: A
authors.lastname: Gulliksen
authors.forename: Helen
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authors.lastname: Keegan
authors.forename: Cara
authors.initials: C
authors.lastname: Martin
authors.forename: John
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authors.lastname: O'Leary
authors.forename: Lars A
authors.initials: LA
authors.lastname: Solli
authors.forename: Inger Marie
authors.initials: IM
authors.lastname: Falang
authors.forename: Petter
authors.initials: P
authors.lastname: Grønn
authors.forename: Aina
authors.initials: A
authors.lastname: Karlgård
authors.forename: Michal M
authors.initials: MM
authors.lastname: Mielnik
authors.forename: Ib-Rune
authors.initials: IR
authors.lastname: Johansen
authors.forename: Terje R
authors.initials: TR
authors.lastname: Tofteberg
authors.forename: Tobias
authors.initials: T
authors.lastname: Baier
authors.forename: Rainer
authors.initials: R
authors.lastname: Gransee
authors.forename: Klaus
authors.initials: K
authors.lastname: Drese
authors.forename: Thomas
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authors.lastname: Hansen-Hagge
authors.forename: Lutz
authors.initials: L
authors.lastname: Riegger
authors.forename: Peter
authors.initials: P
authors.lastname: Koltay
authors.forename: Roland
authors.initials: R
authors.lastname: Zengerle
authors.forename: Frank
authors.initials: F
authors.lastname: Karlsen
authors.forename: Dag
authors.initials: D
authors.lastname: Ausen
authors.forename: Liv
authors.initials: L
authors.lastname: Furuberg