Electrochemical detection of sequence-specific DNA using a DNA probe labeled with aminoferrocene and chitosan modified electrode immobilized with ssDNA

Abstract

The electrochemical detection of sequence-specific DNA using a DNA probe labeled with aminoferrocene (AFC) is reported. Sample ssDNA was immobilized on a chitosan modified glassy carbon electrode. A sequence-known DNA with 256 bp [obtained by polymerase chain reaction (PCR)] was successfully labeled with the electro-active reagent AFC by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide for the first time. This DNA probe labeled with AFC was applied to hybridize with a sequence-unknown DNA sample. Only the complementary sequence (cDNA) could form a double-stranded DNA (dsDNA) with the DNA probe labeled with AFC. The anodic peak currents (ipa) of the AFC bound to the dsDNA by differential pulse voltammetry were used for the determination of cDNA. The ipa of AFC was linearly related to the concentration of cDNA sequence between 1.0 x 10(-8) and 6.0 x 10(-6) mol L-1. The detection limit was 2.0 x 10(-9) mol L-1 using 3 sigma (where sigma is the standard deviation of blank solution, n = 11). The probe showed high sensitivity and selectivity.

Diagnosis of Sugarcane White Leaf Disease Using the Highly Sensitive DNA Based Voltammetric Electrochemical Determination

“…These linear equations might be useful in an analysis for narrowing the actual concentration within each suspected range. The detection capability is comparable to several published electrochemical voltammetric DNA detection such as the detection limit of 5.0 nM via ferrocene labeling and chitosan-modified electrode [33] [34], 0.5 nM for Microcystis related 17 bases DNA sequence immobilized on CPE [22], 0.252 nM of fish sperm DNA via chitosan/CNT/graphite electrode [28], 0.2 nM via a hairpin DNA on gold electrode [29], the detection range of 6.25 -100 nM for 104 nt DNA sequence using PEGylated 20 nt ssDNA and mercaptoethanol on gold electrode [41], 0.1 nM of the synthetic oligonucleotides according to HIV sequence using gold electrode with 6 sensor array [23], the detection range of 10 -60 ng/µl for genomic DNA and PCR product of Neisseria meningitides [42] and the detection limit of 36.72 ng/μL for the concentration range 100 -500 ng/μl of genomic DNA from Vibrio cholera [43]. The results thus demonstrate the great potential for practical application of this electrochemical response detection for the qualitative and quantitative analysis of SCWL with a very simple preparation using domestically available chitosan and self producible DNA probe.…”

Label‐Free and Label Based Electrochemical Detection of Hybridization by Using Methylene Blue and Peptide Nucleic Acid Probes at Chitosan Modified Carbon Paste Electrodes

“…They have detected hybridization by using polymerase chain reaction (PCR) product modified with ferrocenecarboxaldehyde at chitosan modified platinum electrode. Xu et al [38] also reported the detection of hybridization by using aminoferrocene modified oligonucleotides immobilized on glassy carbon electrode.…”

Zinc oxide-chitosan nanobiocomposite for urea sensor

“…Moreover, presence of amine and hydroxyl group in CH helps in the immobilization of desired enzyme. [13][14][15] Recently, ZnO-CH nanobiocomposite films have been proposed for amperometric immunosensor for human IgG. Moreover, ZnO-CH composite film can be used for application to H 2 O 2 , phenol and cholesterol biosensor, respectively.…”