Abstract
Lung cancer is one of the most serious threats to human where 85% of lethal death caused by non-small cell lung cancer (NSCLC) induced by epidermal growth factor receptor (EGFR) mutation. The present research focuses in the development of efficient and effortless EGFR mutant detection strategy through high-performance and sensitive genosensor. The current amplified through 250 µm sized fingers between 100 µm aluminium electrodes indicates the voltammetry signal generated by means of the mutant DNA sequence hybridization. To enhance the DNA immobilization and hybridization, ∼25 nm sized aluminosilicate nanocomposite synthesized from the disposed joss fly ash was deposited on the gaps between aluminium electrodes. The probe, mutant (complementary), and wild (single-base pair mismatch) targets were designed precisely from the genomic sequences denote the detection of EGFR mutation. Fourier-transform Infrared Spectroscopy analysis was performed at every step of surface functionalization evidences the relevant chemical bonding of biomolecules on the genosensor as duplex DNA with peak response at 1150 cm−1 to 1650 cm−1. Genosensor depicts a sensitive EGFR mutation as it is able to detect apparently at 100 aM mutant against 1 µM DNA probe. The insignificant voltammetry signal generated with wild type strand emphasizes the specificity of genosensor in the detection of single base pair mismatch. The inefficiency of genosensor in detecting EGFR mutation in the absence of aluminosilicate nanocomposite implies the insensitivity of genosensing DNA hybridization and accentuates the significance of aluminosilicate. Based on the slope of the calibration curve, the attained sensitivity of aluminosilicate modified genosensor was 3.02E-4 A M−1. The detection limit of genosensor computed based on 3σ calculation, relative to the change of current proportional to the logarithm of mutant concentration is at 100 aM.
Highlights
In the last decade, the application of biosensors in the detection of blood biomarkers, and proteins have received high attention in the diagnosis of various diseases[18,19]
The genosensor assembled in the present research was validated using complementary mutant DNA strand and one base mismatch DNA strand in the hybridization against DNA probe indicating the occurrence of non-small cell lung cancer (NSCLC) via the voltammetry signal amplified through the genosensor
The energy disperses spectroscopy obtained under FESEM analysis showed the ratio of silica to aluminium was 13.24 to 7.96 (1.66) justifies the purity of aluminosilicate nanocomposite synthesized from the environmental waste substances
Summary
The application of biosensors in the detection of blood biomarkers, and proteins have received high attention in the diagnosis of various diseases[18,19]. The sensitivity in hybridization of genomic DNA strands designed for the detection of EGFR mutation indicated by the current conducted through the electrodes of the genosensor.
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