In-Check system: A highly integrated silicon Lab-on-Chip for sample preparation, PCR amplification and microarray detection of nucleic acids directly from biological samples

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• 作者：Salvatore Petralia ; Roberto Verardo ; Enio Klaric ; Sebastiano Cavallaro ; Enrico Alessi ; Claudio Schneider
• 关键词：Lab-on-Chip ; Sample preparation ; PCR (polymerase chain reaction) ; Microarray hybridization
• 刊名：Sensors and Actuators B: Chemical
• 出版年：2013
• 出版时间：October, 2013
• 年：2013
• 卷：187
• 期：Complete
• 页码：99-105
• 全文大小：874 K

文摘

The development of a simple and rapid point-of-care clinical diagnostic device is essential for the evolution of sensing in healthcare technologies. In this study we introduce a fully integrated method for the detection of nucleic acids from cultured human cells and whole blood using the In-Check system supplied by STMicroelectronics.

This work represents the first step toward the integration of sample preparation, nucleic acids amplification and DNA microarray detection into a simple and fast silicon Lab-on-Chip system. In particular, we report the experimental results for the detection of the human ¦Â-globine gene (HBB) directly from defined numbers of cultured human cells and whole blood in less than 2 h. The influence of PCR inhibitors was also evaluated when the detection was performed on whole blood.

The sample preparation process was entirely performed in a single step in the silicon microreactor, and assessed by RT-qPCR and direct DNA quantification. The extracted DNA, after PCR amplification, was hybridized on the DNA microarray. Both the DNA amount and the amplification efficiency were higher than those obtained with a conventional multi-step process used for comparison. Altogether, the results demonstrate the feasibility of casting a sample preparation step in miniaturized microfluidics systems to develop fully integrated, simple and rapid sample-to-detection nucleic acids-based diagnostic systems.