Journal article
Electrochemical detection of leukemia oncogenes using enzyme-loaded carbon nanotube labels
Analyst (London), Vol.139(17), pp.4223-4230
2014
Handle:
https://hdl.handle.net/2376/105711
PMID: 24961450
Abstract
We describe an ultrasensitive electrochemical nucleic acid assay amplified by carbon nanotubes (CNTs)-based labels for the detection of human acute lymphocytic leukemia (ALL)-related p185BCR-ABLfusion transcript. The carboxylated CNTs were functionalized with horseradish peroxidase (HRP) molecules and target-specific detection probes (DP)viadiimide-activated amidation and used to label and amplify the target hybridization signal. The activity of captured HRP was monitored by square-wave voltammetry measuring the electroactive enzymatic product in the presence of 2-aminophenol and hydrogen peroxide substrate solution. The signal-amplified assay achieved a detection limit of 83 fM (5 × 10−18mol in 60 μL) targets oligonucleotides and has a 4-order-wide dynamic range of target concentration. The resulting assay allowed robust discrimination between the perfect match and a three-base mismatch sequence. When exposed to the full-length (491 bp) DNA oncogene, the approach demonstrated a detection limit of 1 × 10−16mol in 60 μL, corresponding to approximately 33 pg of the target gene. The high sensitivity and specificity of the assay enabled a PCR-free detection of target transcripts in as little as 65 ng of mRNA extracted from positive ALL cell lines SUP-B15 in comparison to those obtained from negative cell line HL-60. The approach enables a simple, low-cost and ultrasensitive electrochemical nucleic acid detection in portable devices, point-of-care and early disease diagnostic applications.
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Details
- Title
- Electrochemical detection of leukemia oncogenes using enzyme-loaded carbon nanotube labels
- Creators
- Ai-Cheng Lee - Pacific Northwest National Laboratory, Richland, USA, Department of Biological Sciences, National University of Singapore, , Singapore 117543Dan Du - Pacific Northwest National Laboratory, Richland, USA, School of Mechanical and Materials Engineering, Washington State University, Pullman, USABaowei Chen - Pacific Northwest National Laboratory, Richland, USAChew-Kiat Heng - Department of Paediatrics, National University of Singapore, , SingaporeTit-Meng Lim - Department of Biological Sciences, National University of Singapore, , Singapore 117543Yuehe Lin - Pacific Northwest National Laboratory, Richland, USA, School of Mechanical and Materials Engineering, Washington State University, Pullman, USA
- Publication Details
- Analyst (London), Vol.139(17), pp.4223-4230
- Academic Unit
- School of Mechanical and Materials Engineering
- Identifiers
- 99900546771001842
- Language
- English
- Resource Type
- Journal article