A Nano-Biosensor for the Detection of 185delAG Mutation in BRCA1 Gene, Leading to Breast Cancer (original) (raw)
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This paper demonstrates a simple, label-free detection methodology for detecting single point DNA mutations. Single point mutation detection is a key enabler for diagnosis and prevention of several genetic disorders that manifest into cancers. Specifically for this purpose, herein, an electrochemical biosensor utilizing electrospun graphene doped manganese III oxide nanofibers (GMnO) is developed. The charge transfer resistance offered by GMnO is extremely sensitive to the localized change in the conductivity. This sensitivity, attributed to the low band gap of Mn 2 O 3 and high charge transfer kinetics of graphene, is explored in the proposed mutation detection platform. As a proof of concept, ultrasensitive detection of BRCA1 gene specific point mutation is demonstrated. The target specific single stranded probe DNA is immobilized onto GMnO modified glassy carbon working electrodes via chemisorption. Post target-DNA hybridization, differential pulse voltammetry is employed to facilitate detection of targeted point mutation, wherein, difference in peak currents is used to distinguish the target DNA as normal or mutant. Efficiency of the proposed method is evaluated against a target concentration ranging from 10 pMÀ1 mM. With respect to the mutated target DNA, the LoD of the proposed device is found to be 0.8 AE 0.069 pM. The proposed approach can be extended for detecting any mutation/hybridization of interest by simply adapting an appropriate functionalization protocol. DNA Mutation BRCA1 Differential Pulse Voltammetry Label-free Electrospinning
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In the past few years, graphene oxide (GO) has emerged as a unique platform for developing DNAbased biosensors, given the DNA adsorption and fluorescence-quenching properties of GO. Adsorbed DNA probes can be desorbed from the GO surface in the presence of target analytes, producing a fluorescence signal. In addition to this initial design, many other strategies have been reported, including the use of aptamers, molecular beacons, and DNAzymes as probes, label-free detection, utilization of the intrinsic fluorescence of GO, and the application of covalently linked DNA probes. The potential applications of DNAfunctionalized GO range from environmental monitoring and cell imaging to biomedical diagnosis. In this review, we first summarize the fundamental surface interactions between DNA and GO and the related fluorescencequenching mechanism. Following that, the various sensor design strategies are critically compared. Problems that must be overcome before this technology can reach its full potential are described, and a few future directions are also discussed.
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A numerical illustration of a hybrid design and numerical analysis of graphene-coated fiber-optic surface plasmon resonance (SPR) biosensor for BRCA-1 and BRCA-2 genetic breast cancer detection is provided. Two specific mutations named 916delTT in BRCA-1 gene and 6174delT in BRCA-2 gene are being selected for detection of breast cancer numerically. This sensor is based on attenuated total reflection (ATR) method to detect individual point mutations in BRCA-1 and BRCA-2 genes. Based on the numerically obtained results, a momentous change is present in the SPR angle (minimum 35% more) and surface resonance frequency (SRF) (minimum 36% more) for probe DNA with various concentrations of target DNA corresponding to the mutation of the BRCA-1 and BRCA-2 genes. The variation of the SPR angle and SRF for mismatched DNA strands in BRCA-1 and BRCA-2 genes is quite negligible, whereas that for complementary DNA strands is considerable. This considerable change is essential for proper detection of genetic biomarkers (916delTT and 6174delT) for early breast cancer. To the best of our knowledge, this is the first demonstration of such an adept biosensor for detecting BRCA1 and BRCA2 genetic breast cancer. Here, we used graphene as bimolecular acknowledgement element for improving sensor performance. At the end of the article, the performance in terms of sensitivity is analyzed. Therefore, the proposed biosensor opens a window toward detection of early detection of BRCA-1 and BRCA-2 genetic breast cancers.
Photonic Sensors, 2019
This paper provides a simple hybrid design and numerical analysis of the graphene-coated fiber-optic surface plasmon resonance (SPR) biosensor for breast cancer gene-1 early onset (BRCA1) and breast cancer gene-2 early onset (BRCA2) genetic breast cancer detection. Two specific mutations named 916delTT and 6174delT in the BRCA1 and BRCA2 are selected for numerical detection of breast cancer. This sensor is based on the technique of the attenuated total reflection (ATR) method to detect deoxyribonucleic acid (DNA) hybridization along with individual point mutations in BRCA1 and BRCA2 genes. We have numerically shown that momentous changes present in the SPR angle (minimum: 135 % more) and surface resonance frequency (SRF) (minimum: 136 % more) for probe DNA with various concentrations of target DNA corresponding to a mutation of the BRCA1 and BRCA2 genes. The variation of the SPR angle and SRF for mismatched DNA strands is quite negligible, whereas that for complementary DNA strands is considerable, which is essential for proper detection of genetic biomarkers (916delTT and 6174delT) for early breast cancer. At last, the effect of electric field distribution in inserting graphene layer is analyzed incorporating the finite difference time domain (FDTD) technique by using Lumerical FDTD solution commercial software. To the best of our knowledge, this is the first demonstration of such a highly efficient biosensor for detecting BRCA1 and BRCA2 breast cancer. Therefore, the proposed biosensor opens a new window toward the detection of breast cancers.
Acta chimica Slovenica, 2018
In this study, a simple and novel electrochemical biosensor based on a glassy carbon electrode (GCE) modified with a composite of graphene oxide (GO) - silk fibroin nanofibers (SF) and gold nanoparticles (MCH/ssDNA/AuNPs/SF/GO/GCE) was developed for detection of DNA sequences. The fabrication processes of electrochemical biosensor were characterized by scanning electron microscopy (SEM), FT-IR and electrochemical methods. Some experimental conditions such as immobilization time of probe DNA and MCH incubation time, time and temperature of hybridization were optimized. The designed biosensor revealed a wide linear range of 1.0 × 10-16 - 1.0 × 10-8 mol L-1 and a low detection limit (3.3 × 10-17 mol L-1) for detection of BRCA1 5382 mutation by EIS technique. The designed biosensor revealed high selectivity for discrimination of the complementary (P1C) sequences from various non-complementary sequences of (P1nC1, P1nC2 and P1nC3). Also, the biosensor revealed a high reproducibility (RSD...
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