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Journal of Oncology
Volume 2012, Article ID 905024, 12 pages
http://dx.doi.org/10.1155/2012/905024
Research Article

Towards a “Sample-In, Answer-Out” Point-of-Care Platform for Nucleic Acid Extraction and Amplification: Using an HPV E6/E7 mRNA Model System

1NorChip AS, Industriveien 8, 3490 Klokkarstua, Norway
2University of Oslo, 0316 Oslo, Norway
3Department of Histopathology, Trinity College Dublin and Molecular Pathology Research Laboratory, Coombe Women and Infants University Hospital, Dolphins Barn, Dublin 8, Ireland
4SINTEF ICT, MiNaLab Facility, Gaustadalléen 23C, 0373 Oslo, Norway
5Institut für Mikrotechnik Mainz, Carl-Zeiss Straße 18-20, 55129 Mainz, Germany
6Center of Smart Interfaces, TU Darmstadt, Petersenstraße 32, 64287 Darmstadt, Germany
7Laboratory for MEMS Applications, Department of Microsystems Engineering-IMTEK, University of Freiburg, Georges-Koehler-Allee 106, 79110 Freiburg, Germany
8BioFluidix GmbH, Georges-Koehler-Allee 106, 79110 Freiburg, Germany
9Department of Micro and Nano Systems Technology, Faculty of Technology and Maritime Sciences, Vestfold University College, Raveien 197, 3184 Borre, Norway

Received 21 July 2011; Accepted 6 September 2011

Academic Editor: Adhemar Longatto-Filho

Copyright © 2012 Anja Gulliksen et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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 = 2 8 ) 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.