Anja Gulliksen,
Helen Keegan,
Cara Martin,
John O'Leary,
Lars A Solli,
Inger Marie Falang,
Petter Grønn,
Aina Karlgård,
Michal M Mielnik,
Ib-Rune Johansen, [......],
Tobias Baier,
Rainer Gransee,
Klaus Drese, Thomas Hansen-Hagge,
Lutz Riegger,
Peter Koltay,
Roland Zengerle,
Frank Karlsen,
Dag Ausen,
Liv Furuberg
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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.
Journal of Oncology 01/2012; 2012:905024.
