μ-eLCR: a microfabricated device for electrochemical detection of DNA base changes in breast cancer cell lines.
Microfabricated devices for the electrochemical detection of single DNA base changes in cancer cell lines are highly desirable due to the inherent advantages such as portability, simplicity, and the rapid and inexpensive nature of electrochemical readout methods. Moreover, molecular sensors that use microscale-footprint working electrodes have shown high signal-to-noise ratio. Herein we report a microdevice-based electrochemical assay (μ-eLCR) measuring ligase chain reaction (LCR)-amplified long and short "knife" motifs which reflect the presence or absence of a DNA base change of interest in a target sequence. This μ-eLCR approach has higher sensitivity (4.4 to 10 fold improvement over macrodisk electrodes) and good reproducibility (%RSD 6.5%, n = 12) for the detection of LCR-amplified DNA bases. The devices also exhibited excellent sensitivity for the detection of DNA methylation (C to T base change in a locus associated with cancer metastasis) in two cell lines and serum derived DNA samples. We believe that the μ-eLCR device may be a useful diagnostic tool for inexpensive and rapid detection of single DNA base change applications such as DNA methylation and single nucleotide polymorphism (SNP) detection.
Centre for Biomarker Research and Development, Australian Institute for Bioengineering and Nanotechnology (AIBN), Brisbane, QLD 4072, Australia. m.shiddiky@uq.edu.au.
Eugene J H
EJ
Wee
Sakandar
S
Rauf
Kevin M
KM
Koo
Muhammad J A
MJ
Shiddiky
Matt
M
Trau
D001483:Base Sequence; D001943:Breast Neoplasms / Q000235:genetics; D045744:Cell Line, Tumor; D019175:DNA Methylation; D015342:DNA Probes / Q000737:chemistry / Q000378:metabolism; D004273:DNA, Neoplasm / Q000032:analysis*; D055664:Electrochemical Techniques / Q000295:instrumentation / Q000379:methods*; D004566:Electrodes; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry; D006046:Gold / Q000737:chemistry; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D021141:Nucleic Acid Amplification Techniques; D020641:Polymorphism, Single Nucleotide
D015342:DNA Probes; D004273:DNA, Neoplasm; D005456:Fluorescent Dyes; D006046:Gold