On-chip LAMP-BART reaction for viral DNA real-time bioluminescence detection

• Published in: Sensors and actuators. B, Chemical Vol. 262; pp. 1024 - 1033
• Main Authors: Mirasoli, M., Bonvicini, F., Lovecchio, N., Petrucci, G., Zangheri, M., Calabria, D., Costantini, F., Roda, A., Gallinella, G., Caputo, D., de Cesare, G., Nascetti, A.
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 01-06-2018; Elsevier Science Ltd
• Subjects: Amorphous silicon; Amorphous silicon sensors; Bioluminescence; Bioluminescent assay in real time BART; Deoxyribonucleic acid; DNA; Glass substrates; Lab-on-a-chip; Lab-on-chip; Loop-mediated isothermal amplification LAMP; Polydimethylsiloxane; Radiation detectors; Real time; Sensors; Silicon diodes; Studies; Target detection; Thin film technology; Thin films; Viral nucleic acids detection; Lab-on-chip; Amorphous silicon sensors; Bioluminescent assay in real time BART; Thin film technology; Loop-mediated isothermal amplification LAMP; Viral nucleic acids detection
• ISSN: 0925-4005; 1873-3077
• DOI: 10.1016/j.snb.2018.02.086

•Development and testing of a lab-on-chip for viral DNA amplification with real-time on-chip detection.•Optimization of a loop-mediated isothermal amplification (LAMP) technique to specifically amplify parvovirus B19 DNA.•Coupling of LAMP with Bioluminescent Assay in Real Time (BART) technology to provide real-time quantification of target DNA.•Integration of amorphous silicon sensors and thin film heaters on a glass substrate for temperature control and on-chip detection.
The present paper describes the development of an integrated lab-on-chip, in which viral DNA amplification with real-time on-chip detection is carried out under constant temperature of 65 °C. The lab-on-chip is composed of a disposable 10-μL polydimethylsiloxane reaction chamber which is thermally and optically coupled to a glass substrate that hosts a thin-film metallic resistive heater and thin-film amorphous silicon diodes which act as temperature and radiation sensors. A loop-mediated isothermal amplification (LAMP) technique was optimized to specifically amplify parvovirus B19 DNA and coupled with Bioluminescent Assay in Real Time (BART) technology to provide real-time detection of target DNA. The experimental results demonstrate the ability of the proposed device to discriminate among different concentrations of viral DNA with an excellent agreement with standard off-chip methods.