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- Title
Simultaneous DNA amplification and detection using a pH-sensing semiconductor system.
- Authors
Toumazou, Christofer; Shepherd, Leila M; Reed, Samuel C; Chen, Ginny I; Patel, Alpesh; Garner, David M; Wang, Chan-Ju A; Ou, Chung-Pei; Amin-Desai, Krishna; Athanasiou, Panteleimon; Bai, Hua; Brizido, Ines M Q; Caldwell, Benjamin; Coomber-Alford, Daniel; Georgiou, Pantelis; Jordan, Karen S; Joyce, John C; La Mura, Maurizio; Morley, Daniel; Sathyavruthan, Sreekala; Temelso, Sara; Thomas, Risha E; Zhang, Linglan
- Abstract
We developed an integrated chip for real-time amplification and detection of nucleic acid using pH-sensing complementary metal-oxide semiconductor (CMOS) technology. Here we show an amplification-coupled detection method for directly measuring released hydrogen ions during nucleotide incorporation rather than relying on indirect measurements such as fluorescent dyes. This is a label-free, non-optical, real-time method for detecting and quantifying target sequences by monitoring pH signatures of native amplification chemistries. The chip has ion-sensitive field effect transistor (ISFET) sensors, temperature sensors, resistive heating, signal processing and control circuitry all integrated to create a full system-on-chip platform. We evaluated the platform using two amplification strategies: PCR and isothermal amplification. Using this platform, we genotyped and discriminated unique single-nucleotide polymorphism (SNP) variants of the cytochrome P450 family from crude human saliva. We anticipate this semiconductor technology will enable the creation of devices for cost-effective, portable and scalable real-time nucleic acid analysis.
- Publication
Nature methods, 2013, Vol 10, Issue 7, p641
- ISSN
1548-7105
- Publication type
Journal Article
- DOI
10.1038/nmeth.2520