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Rapid nested-PCR for tyrosinase gene detection on chip
journal contributionposted on 11.06.2015, 15:27 by Anna G. Sciancalepore, Alessandro Polini, Elisa MeleElisa Mele, Salvatore Girardo, Roberto Cingolani, Dario Pisignano
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.
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.
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