ИСТИНА

Development of new express diagnostic tools is very important for the future medicine, agriculture and environmental science. Key components of such systems are usually chips with different levels of complexity. The mainstream of chip development is miniaturisation of widely used laboratory methods like PCR, hybridization and immunoassay which provide bio-chemical detection basement. Miniaturisation opens up new possibilities like shortening time to result, makes operating device simpler and cheaper. At the same time, chips are usually compromise sensitivity because of lack of proper sample preparation and reduction of sample volume. Another problem of the chip systems is limited multiplexing. PCR on the chip is very perspective because PCR by itself is the most sensitive among biochemical methods and with new chemistries does not require complex sample preparation procedures. In frame of the present work, we tried to overcome common problems of the chip-PCR. PDMS, PC and Zeonor were used to develop chip-PCR. Real-time in rhombical chip and continuous flow PCR were developed. Integrations of PCR modules present on Fig 1. Continuous flow PCR-chip was thermocycled on plate with two constant zones (64°C and 96°C) (Fig. 1A) while rhombical chip with Peltier element (Fig. 1B). Both systems were capable to produce PCR products in single- and multiplex formats with sensitivity around 10 3 CFU/ml for less than an hour. Escherichia coli, Salmonella typhi and Mycobacterium smegmatis were used as controls for experimental detection. Best results were produces with PC and Zeonor chips. Both chip variants will be used to develop system with automatic chip replacement and multiplex detection for up to 30 targets on the chip.