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Title: Rapid nested-PCR for tyrosinase gene detection on chip
Authors: Sciancalepore, Anna G.
Polini, Alessandro
Mele, Elisa
Girardo, Salvatore
Cingolani, Roberto
Pisignano, Dario
Keywords: Tyrosinase
Issue Date: 2011
Publisher: © Elsevier
Citation: SCIANCALEPORE, A.G. ... et al, 2011. Rapid nested-PCR for tyrosinase gene detection on chip. Biosensors and Bioelectronics, 26 (5), pp. 2711 - 2715.
Abstract: The availability of non-invasive, fast and sensitive technologies for detection of circulating cancer cells is still a critical need of clinical oncology, particularly for diagnosis of aggressive and highly metastatic tumors, like malignant melanoma. Here we present the first nested polymerase chain reaction process carried out by a microfabricated, hybrid plastic–glass microfluidic chip on the tyrosinase gene, a predictive marker for melanoma diagnosis. The device is a hybrid system consisting of a glass microchannel embedded in an elastomeric matrix, and operating in flow-oscillating modality on a droplet of biological sample. The convection heat transfer and the temperature distribution inside the carrier fluid in the device are investigated. The oil responds to temperature changes with a characteristic time around 53 s, and exhibits three different thermal gradients along the capillary, with temperature variations below 4 ◦C in correspondence of heater electrodes. The sample heating/cooling rates in the chip are as high as 16 ◦C/s, allowing rapid processes. The nested polymerase chain reaction process is performed in less than 50 min, namely more than four times faster than in a standard thermocycler. The rapidity of the analysis method, combined with the simple and low-cost fabrication, reduced sample evaporation, and flexibility of the overall microfluidic platform, make it promising for the detection of events of tumor spreading.
Description: This article is closed access.
Sponsor: The authors are grateful to the support of the Apulia Regional Strategic Project PS 144, and of the Italian Ministry of University and Research through the FIRB project RBLA03ER38.
Version: Published
DOI: 10.1016/j.bios.2010.09.008
URI: https://dspace.lboro.ac.uk/2134/17817
Publisher Link: http://dx.doi.org/10.1016/j.bios.2010.09.008
ISSN: 0956-5663
Appears in Collections:Closed Access (Materials)

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