Microchip analytic system for multiplex analysis by real-time polymerase chain reaction with reagents immobilized in microreactors

A microchip analytic system that uses a silicon chip with immobilized in microreactor test-system for multiplex analysis of DNA by real-time polymerase chain reaction (real time PCR) was developed and optimized. We suggested the method of immobilization of PCR-components of a test-system, chose the stabilizer, and conducted the optimization of the composition of reaction mixture to achieve permanent stability of a microchip. We conducted optimization of preparation of samples using magnetic sorbent and indicated that, with 2.6 × 10⁴ copies/ml, 60 min are necessary to obtain positive identification including time for preparation of model samples. The abilities of the created system were demonstrated on the example of microchip analysis of samples with different content of DNA, low absolute limits of detection (20 DNA copies in microreactor), and high reproducibility of the analysis.