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

Autor: Tobias Baier, Roland Zengerle, Liv Furuberg, Klaus Stefan Drese, Michal Marek Mielnik, Peter Koltay, Aina Karlgård, Dag Ausen, Lars Anders Solli, John J. O'Leary, Rainer Gransee, Ib-Rune Johansen, Thomas Hansen-Hagge, Cara Martin, Frank Karlsen, Petter Grønn, Helen Keegan, L. Riegger, Terje Rosquist Tofteberg, Inger Marie Falang, Anja Gulliksen
Jazyk: angličtina
Rok vydání: 2012
Předmět:
Zdroj: Journal of Oncology
Journal of Oncology, Vol 2012 (2012)
ISSN: 1687-8450
DOI: 10.1155/2012/905024
Popis: 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.
Databáze: OpenAIRE
Externí odkaz: