On-chip quantitative-PCR using integrated real-time detection by capillary electrophoresis

• Published in: Electrophoresis Vol. 37; no. 3
• Main Authors: Liu, Yu, Li, Chen, Li, Zhi, Chan, Samuel D., Eto, Daisuke, Wu, Warren, Zhang, Jian Ping, Chien, Ring-Ling, Wada, Henry G., Greenstein, Michael, Satomura, Shinji
• Format: Journal Article
• Language: English
• Published: Blackwell Publishing Ltd 01.02.2016
• Subjects: Assaying; Capillarity; Channels; Design analysis; Electrophoresis; Real time; Separation; Synchronism
• ISSN: 0173-0835; 1522-2683
• DOI: 10.1002/elps.201670031

Electrophoresis 2016, 37, 545–552. DOI: 10.1002/elps.201500298 Quantitative polymerase chain reaction (qPCR) and capillary electrophoresis (CE) were integrated into a microfluidic chip that was designed to achieve real‐time amplicon sampling, separation, and quantitation without requiring various probes. A novel channel design and PCR‐CE synchronization scheme allows the overlapped execution of PCR and CE, minimizing the time required for CE analysis after each PCR cycle. The performance of the on‐chip qPCRCE method was demonstrated using model assay protocols to detect phiX174 bacteriophage and E. coli genomic DNA. By combining the separation capability of CE and the amplification power of PCR assay, this method is expected to find applications in awide variety of infectious disease detection and monitoring assays.