title: Simultaneous DNA amplification and detection using a pH-sensing semiconductor system.
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.
authors.affiliation: DNA Electronics Ltd., London, UK. chris.toumazou@dnae.co.uk
authors.forename: Christofer
authors.initials: C
authors.lastname: Toumazou
authors.forename: Leila M
authors.initials: LM
authors.lastname: Shepherd
authors.forename: Samuel C
authors.initials: SC
authors.lastname: Reed
authors.forename: Ginny I
authors.initials: GI
authors.lastname: Chen
authors.forename: Alpesh
authors.initials: A
authors.lastname: Patel
authors.forename: David M
authors.initials: DM
authors.lastname: Garner
authors.forename: Chan-Ju A
authors.initials: CJ
authors.lastname: Wang
authors.forename: Chung-Pei
authors.initials: CP
authors.lastname: Ou
authors.forename: Krishna
authors.initials: K
authors.lastname: Amin-Desai
authors.forename: Panteleimon
authors.initials: P
authors.lastname: Athanasiou
authors.forename: Hua
authors.initials: H
authors.lastname: Bai
authors.forename: Ines M Q
authors.initials: IM
authors.lastname: Brizido
authors.forename: Benjamin
authors.initials: B
authors.lastname: Caldwell
authors.forename: Daniel
authors.initials: D
authors.lastname: Coomber-Alford
authors.forename: Pantelis
authors.initials: P
authors.lastname: Georgiou
authors.forename: Karen S
authors.initials: KS
authors.lastname: Jordan
authors.forename: John C
authors.initials: JC
authors.lastname: Joyce
authors.forename: Maurizio
authors.initials: M
authors.lastname: La Mura
authors.forename: Daniel
authors.initials: D
authors.lastname: Morley
authors.forename: Sreekala
authors.initials: S
authors.lastname: Sathyavruthan
authors.forename: Sara
authors.initials: S
authors.lastname: Temelso
authors.forename: Risha E
authors.initials: RE
authors.lastname: Thomas
authors.forename: Linglan
authors.initials: L
authors.lastname: Zhang
mesh_terms: D004867:Equipment Design; D006863:Hydrogen-Ion Concentration; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation*; D012666:Semiconductors; D017422:Sequence Analysis, DNA / Q000295:instrumentation*; D012815:Signal Processing, Computer-Assisted / Q000295:instrumentation*; D018511:Systems Integration