DNA detection on transistor arrays following mutation-specific enzymatic amplification

Abstract

An integrated array of silicon field-effect transistor structures is used for electronic detection of label-free DNA. Measurements of the dc current–voltage characteristics of the transistors gives us access to reproducible detection of single- and double-stranded DNA, locally adsorbed on the surface of the device. We combine this approach with allele-specific polymerase chain reaction, to test for the 35delG mutation, a frequent mutation related to prelingual nonsyndromic deafness.

Field-effect detection using phospholipid membranes

“…There are various combinations of analytes, recognition elements and semiconductors. Recent examples of detected biological events include protein binding [1][2][3][4][5][6], enzymatic reactions [7], DNA hybridization [8][9][10], DNA or RNA hybridization with peptide nucleic acid (PNA) [11,12], and the binding of other biologically related molecules [13].…”

Real-time label-free quantitative monitoring of biomolecules without surface binding by floating-gate complementary metal-oxide semiconductor sensor array integrated with readout circuitry

“…6,7 However, in order to detect DNA molecules, it is required to immobilize single-stranded probes on the surface of the ion-sensitive devices for the hybridization of single-stranded complementary target molecules in the electrolyte solution [8][9][10] or to deposit a positively charged layer such as poly-L-lysine on the sensing electrodes to attract the negatively charged nucleic acid molecules. 11,12 These additional processes increase reaction time and the complexity in detection protocols. Furthermore, it is difficult to incorporate the continuous monitoring for repetitive assays ͑e.g., real-time quantitative PCR͒ for these protocols because the regeneration of the sensing surface requires a complicated rinsing sequence.…”

Electronic Detection of DNA Adsorption and Hybridization

“…In a first approach, point-mutations have been detected in human DNA by allele-specific PCR followed by electronic detection of the double stranded PCR product [15]. The PCR and the electronic detection are separated by a purification that retains PCR product and substrat DNA, but eliminates nucleotides, primers, proteines and salt.…”

“…In another study, we showed that the FET based measurement is compatible with enzymatic technology and complex DNA samples [15]. We combined the electronic detection with an allele-specific polymerase chain reaction and thus detected a single-basepair mutation in genomic DNA.…”

“…We use a differential technique based on dc measurements with arrays of integrated FET devices [12,15,16]. Our approach allows for efficient miniaturisation and parallelisation through on-chip integration of the detection and readout circuits [17].…”