Label-Free Electrochemical Detection of DNA Hybridization Related to Anthrax Lethal Factor by using Carbon Nanotube Modified Sensors

2019 ◽  
Vol 15 (4) ◽  
pp. 502-510 ◽  
Author(s):  
Hakan Karadeniz ◽  
Arzum Erdem

Background: Anthrax Lethal Factor (ANT) is the dominant virulence factor produced by B. anthracis and is the major cause of death of infected animals. In this paper, pencil graphite electrodes GE were modified with single-walled and multi-walled carbon nanotubes (CNTs) for the detection of hybridization related to the ANT DNA for the first time in the literature. Methods: The electrochemical monitoring of label-free DNA hybridization related to ANT DNA was explored using both SCNT and MCNT modified PGEs with differential pulse voltammetry (DPV). The performance characteristics of ANT-DNA hybridization on disposable GEs were explored by measuring the guanine signal in terms of optimum analytical conditions; the concentration of SCNT and MCNT, the concentrations of probe and target, and also the hybridization time. Under the optimum conditions, the selectivity of probe modified electrodes was tested and the detection limit was calculated. Results: The selectivity of ANT probes immobilized onto MCNT-GEs was tested in the presence of hybridization of probe with NC no response was observed and with MM, smaller responses were observed in comparison to full-match DNA hybridization case. Even though there are unwanted substituents in the mixture samples containing both the target and NC in the ratio 1:1 and both the target and MM in the ratio 1:1, it has been found that ANT probe immobilized CNT modified graphite sensor can also select its target by resulting with 20.9% decreased response in comparison to the one measured in the case of full-match DNA hybridization case Therefore, it was concluded that the detection of direct DNA hybridization was performed by using MCNT-GEs with an acceptable selectivity. Conclusion: Disposable SCNT/MCNT modified GEs bring some important advantages to our assay including easy use, cost-effectiveness and giving a response in a shorter time compared to unmodified PGE, carbon paste electrode and glassy carbon electrode developed for electrochemical monitoring of DNA hybridization. Consequently, the detection of DNA hybridization related to the ANT DNA by MCNT modified sensors was performed by using lower CNT, probe and target concentrations, in a shorter hybridization time and resulting in a lower detection limit according to the SCNT modified sensors. In conclusion, MCNT modified sensors can yield the possibilities leading to the development of nucleic acid sensors platforms for the improvement of fast and cost-effective detection systems with respect to DNA chip technology.

2004 ◽  
Vol 16 (11) ◽  
pp. 928-931 ◽  
Author(s):  
Joseph Wang ◽  
Gerd-Uwe Flechsig ◽  
Arzum Erdem ◽  
Olga Korbut ◽  
Peter Gründler

Talanta ◽  
2010 ◽  
Vol 82 (5) ◽  
pp. 1680-1686 ◽  
Author(s):  
Mihrican Muti ◽  
Filiz Kuralay ◽  
Arzum Erdem ◽  
Serdar Abaci ◽  
Tugrul Yumak ◽  
...  

Biosensors ◽  
2022 ◽  
Vol 12 (1) ◽  
pp. 39
Author(s):  
Elena A. Chiticaru ◽  
Luisa Pilan ◽  
Mariana Ioniţă

In this paper, we propose an improved electrochemical platform based on graphene for the detection of DNA hybridization. Commercial screen-printed carbon electrodes (SPCEs) were used for this purpose due to their ease of functionalization and miniaturization opportunities. SPCEs were modified with reduced graphene oxide (RGO), offering a suitable surface for further functionalization. Therefore, aryl-carboxyl groups were integrated onto RGO-modified electrodes by electrochemical reduction of the corresponding diazonium salt to provide enough reaction sites for the covalent immobilization of amino-modified DNA probes. Our final goal was to determine the optimum conditions needed to fabricate a simple, label-free RGO-based electrochemical platform to detect the hybridization between two complementary single-stranded DNA molecules. Each modification step in the fabrication process was monitored by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) using [Fe(CN)6]3−/4− as a redox reporter. Although, the diazonium electrografted layer displayed the expected blocking effect of the charge transfer, the next steps in the modification procedure resulted in enhanced electron transfer properties of the electrode interface. We suggest that the improvement in the charge transfer after the DNA hybridization process could be exploited as a prospective sensing feature. The morphological and structural characterization of the modified electrodes performed by scanning electron microscopy (SEM) and Raman spectroscopy, respectively, were used to validate different modification steps in the platform fabrication process.


2019 ◽  
Vol 9 (3) ◽  
pp. 408-413 ◽  
Author(s):  
Amir H. Keihan ◽  
Ghader Hosseinzadeh ◽  
Sharareh Sajjadi ◽  
Danial Ashiani ◽  
Fariba Dashtestani ◽  
...  

Background: Escherichia coli (E. coli) bacteria is one of the hazardous human pathogens. Consequently, developing the rapid and effective method for identification and quantization of E. coli is popular in biotechnological researches in recent years. Experimental: In this research, a label-free capacitance E. coli biosensor was fabricated based on immobilizing bacteriophage on the carbon paste electrode (Cp). Reduced graphene (RGr) was synthesized and used as a substrate for immobilization of bacteriophage on the Cp surface. E. coli bacteriophage was trapped in graphene modified carbon paste electrodes. The immobilization accuracy was confirmed via electrochemical techniques. The modified electrodes were applied as indicator electrodes for capacitance measurements of E. coli. Results: Through this method, E. coli was detected in a concentration range of 33×10-3 to 330×10-3 N L-1 (number of E. coli per Liter) with a correlation coefficient of 0.99 and a detection limit of 12×10-3 N L-1. Conclusion: The proposed biosensor has a fast response time of about 5 s and good selectivity over other bacteria.


2014 ◽  
Vol 6 (10) ◽  
pp. 7334-7339 ◽  
Author(s):  
Liang Wang ◽  
Yujing Han ◽  
Shuo Zhou ◽  
Guihua Wang ◽  
Xiyun Guan

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Rozita Abolhasan ◽  
Balal Khalilzadeh ◽  
Hadi Yousefi ◽  
Sahar Samemaleki ◽  
Forough Chakari-Khiavi ◽  
...  

AbstractIn the present article, we developed a highly sensitive label-free electrochemical immunosensor based on NiFe-layered double hydroxides (LDH)/reduced graphene oxide (rGO)/gold nanoparticles modified glassy carbon electrode for the determination of receptor tyrosine kinase-like orphan receptor (ROR)-1. In this electrochemical immunoassay platform, NiFe-LDH/rGO was used due to great electron mobility, high specific surface area and flexible structures, while Au nanoparticles were prepared and coated on the modified electrodes to improve the detection sensitivity and ROR1 antibody immobilizing (ROR1Ab). The modification procedure was approved by using cyclic voltammetry and differential pulse voltammetry based on the response of peak current to the step by step modifications. Under optimum conditions, the experimental results showed that the immunosensor revealed a sensitive response to ROR1 in the range of 0.01–1 pg mL−1, and with a lower limit of quantification of 10 attogram/mL (10 ag mL−1). Furthermore, the designed immunosensor was applied for the analysis of ROR1 in several serum samples of chronic lymphocytic leukemia suffering patients with acceptable results, and it also exhibited good selectivity, reproducibility and stability.


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