Single step cell lysis/PCR detection of Escherichia coli in an independently controllable silicon microreactor

Our studies describe a novel microreactor capable of single step microbial assays involving cell lysis and DNA amplification. The device with an integrated platinum heater and temperature sensor, was fabricated using conventional silicon fabrication technologies and then anodically bonded to a Pyrex lid. Finite element analysis (FEA) and experiments have shown that the temperature uniformity in the microreactor reaction cavity is homogeneous and that the microreactor is capable of fast thermal cycling with heating and cooling rates of 11 and 2.7 degrees C/s, respectively. The microreactor has novel design features, such as a thermal isolation channel which eliminates thermal cross talk and an inlet/outlet port designed for ease of use. The fabricated microreactor was successfully characterised using a multifunction microbial assay involving cell lysis and PCR in a single step. An assay time of 32 min was achieved. (c) 2006 Elsevier B.V. All rights reserved.