Array Chip Real Time Quantitative PCR For Gene Expression Analysis

With the rapid development of modern medicine and molecular biology, real-time quantitative PCR technology has played an increasing role both in basic reaserch and clinical tests. In this paper, with a home made static array chip PCR systems and a laser-induced fluorescence detection platform, CXCR-1gene expression analysis was explored, and the applicability of the system for clinical analysis studies was demonstrated.In the first chapter the molecular basis of genetic engineering and the significance of analysis CXCR-1were introduced, and the development of quantitative PCR, especially that based on microfluidic and chip technolobies was reviewed.In the second chapter, the chip system with a diode laser(487/532nm) based laser-induced fluorescence (LIF) detection system was used in the real time PCR, and the conditions for CXCR-1analysis was optimized.In the third chapter, under optimized conditions, quantitative PCR of CXCR-1was performed, and the calibration curves were obtained for the CXCR-1from two clinical samples, and the results were compared with that obtained by a desktop commercial system with good agreement. The chip system showed a better sensitivity and the time is about two times reduced. Parallel real time detection was studied by fluorescent imaging of the array chip, preliminary results gave promising results.This study shows that the chip system is potentially useful in clinics, and the throughput could be greatly increased by parallel detection. We envision that the chip system could be developed into a portable system and may find applications in point of care tests(POCT) in the future.