Lih Poh Lin
ABSTRACT
In this study, a specific impedimetric DNA sensor based on graphene/zirconia nanocomposite has been developed to serve as a simple and rapid method for epidermal growth factor receptor (EGFR) mutations testing, necessary to guide the treatment regime of tyrosine kinase inhibitors (TKIs) therapy. The graphene/zirconia nanocomposite was synthesized via a green method without the use of harsh chemicals. The as-synthesized nanocomposite was characterized with scanning electron microscopy (SEM), X-ray diffractometry (XRD) and energy dispersive X-ray spectroscopy (EDX) to evaluate its morphology, crystallinity and composition. The nanocomposite was functionalized with 1-pyrenebutyric acid N-hydroxysuccinimide ester (PSE) and dropped casted on screen printed electrode (SPCE) to provide amide bonding with single-stranded probe-DNA that is complementary to exon-19 wild-type DNA sequence. Electrochemical responses of each assembly step of the sensing platform were characterized by electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV). Deposition of PSE-functionalized graphene/zirconia and immobilization of single-stranded DNA on the electrode were reflected by the increment of electron transfer resistance (Rct) in EIS analysis as well as the reduction of peak current (ip) in CV analysis. The modified electrodes were hybridized with different target DNAs and the difference in electrochemical impedance before and after hybridization (normalized value) was adopted as the measurement for label-free DNA hybridization detection. The developed genosensor demonstrated strong distinction between the Rct for exon-19 wild-type DNA to exon-19 mutated DNA, leading to promising quantitation of DNA mutations. The biosensor also exhibited high specificity to exon-19 sequence with clear discrimination against non-complementary target. The developed biosensor is disposable and requires no labelling of probe or target, making it beneficial in terms of simplicity and efficiency.
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Index Terms
- Electrochemical DNA Sensor Based On Graphene/Zirconia Nanocomposite for Label-Free Detection of Exon-19 Mutations in Lung Cancer: A Preliminary Study
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