Electrochemical DNA biosensor for the detection of human papillomavirus E6 gene inserted in recombinant plasmid

Abstract

In the current study, we describe a novel, simple, inexpensive, sensitive, specific, stable and label-free electrochemical DNA biosensor used to identify a target gene cloned into a plasmid. The biosensor was designed with a 23-mer oligonucleotide of guanine-free, which was immobilized on the pencil graphite electrode (PGE) for E6 gene detection from human papillomavirus 16 type (HPV16). The E6 gene was used due to its clinical importance. The optimal probe concentration was obtained in 500 nM. The hybridization detection showed a good linearity in the range of 40-5,000 pg/lL with a detection limit of 16 pg/lL. The electrochemical method showed higher sensitivity and specificity when compared with the agarose gel electrophoresis assay. This technology could be postulated as a new and attractive alternative for cloning analysis in plasmids.ª 2014 Production and hosting by Elsevier B.V. on behalf of King Saud University.

A Sensitive and Selective Label-Free Electrochemical DNA Biosensor for the Detection of Specific Dengue Virus Serotype 3 Sequences

“…Finally, a 22-m oligonucleotide was selected to detect DENV-3, with the following sequence: 5′-TAA CAT CAT CAT GAG ACA GAG C-3′. This sequence was selected due to its suitable features that are desirable for electrochemical biosensors, such as shorter base pair length, high specificity and a considerable difference in the number of guanine base between the probe and target sequences, which is important to discriminate between ssDNA and dsDNA onto the PGE surface [ 33 , 39 , 40 ].…”

“…The results showed that the current peaks increase with the increasing concentration of the target sequence (10 nM to 500 nM); the highest concentration exhibited the highest current peak of the system (135 ± 2.15 nA). However, at concentrations higher than 500 nM, there is a decrease in the electrochemical signal that could be due to electrostatic hindrance of DNA molecules on the PGE surface [ 39 , 53 , 56 ].…”

Specific Magnetic Isolation of E6 HPV16 Modified Magnetizable Particles Coupled with PCR and Electrochemical Detection

“…Wide range of electrochemical techniques has been used for this aim such as DPV, SWV, AdTS SWV, CV, etc. [ 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 ].…”

Trends in Biosensors for HPV: Identification and Diagnosis

“…These label-free platforms are developed through the immobilization of the biorecognition probe atop of an interdigitated gold or platinum electrode. The signal transducers are widely diverse, pencil graphite surfaces [30,33], carbon surfaces modified with linear polysaccharides [35], plain glassy carbon surface [34], or poly(methylmethacrylate) substrate recovered with a thin gold film [38]. Additionally, more sophisticated nanostructured ones as graphene/gold nanoparticles (AuNPs), nanorod/poly(thionine), or polyaniline-multiwall carbon nanotube (MWCNT) film [39,68] have been used to construct biosensors committed to detect and differentiate HPV strains.…”

“…Additionally, more sophisticated nanostructured ones as graphene/gold nanoparticles (AuNPs), nanorod/poly(thionine), or polyaniline-multiwall carbon nanotube (MWCNT) film [39,68] have been used to construct biosensors committed to detect and differentiate HPV strains. These platforms facilitate the study of the output signal by cyclic voltammetry (CV), square wave voltammetry (SWV) [33,35], differential pulse voltammetry (DPV) [30,68], and electrochemical impedance spectroscopy (EIS) [68].…”