An Electrochemical Biosensor for the Determination ofGanoderma boninensePathogen Based on a Novel Modified Gold Nanocomposite Film Electrode

Abstract

A sensitive approach for the determination of Ganoderma boninense DNA is reported based on an electrochemical affinity system using a modified gold sensor. Covalent attachment of probe DNA was achieved by attachment of the amine group to a carboxylic acid group of a 3,3'-dithiodipropionic acid monolayer on a nanocomposite film of gold nanoparticles bound to poly(3,4-ethylenedioxythiophen)-poly(styrenesulfonate) on a gold working electrode. The electrochemical detection of sequence-specific DNA of probe and target DNA hybridization was monitored using a new ruthenium complex [Ru(dppz) 2 (qtpy)Cl 2 ; dppz = dipyrido [3,2-a:2',3'-c] phenazine; qtpy = 2,2',-4,4".4'4"'-quarterpyridyl redox marker. The potential was selected through the study of the electrochemical behavior of trisaminomethane-hydrochloride containing a ethylenediaminetetraacetic acid supporting electrolyte on the bare and modified gold electrode. The effect of the hybridization temperature and time were measured. The sensor demonstrated specific detection for the target over a concentration range of 1.0 × 10 −15 M to 1.0 × 10 −9 M with a detection limit of 1.59 × 10 −17 M. Control experiments verified the specificity of the biosensor in the presence of a single mismatched DNA sequence. This detection technology was shown to be effective in terms of sensitivity and selectivity of hybridization events and is a promising device for early detection of Ganoderma boninense and other pathogenic threat agents.

Scientific Importance of Water‐Processable PEDOT–PSS and Preparation, Challenge and New Application in Sensors of Its Film Electrode: A Review

“…Noguer et al designed PEDOT–PSS screen‐printed biosensor based on tyrosinase or acetylcholinesterase for the detection of chlorpyrifos‐oxon and phenolic compounds, respectively . Dutse et al drop‐coated PEDOT–PSS‐AgNPs onto gold electrode for fabricating DNA‐based biosensor, and ganoderma boninense was successfully detected using ruthenium complex [Ru(phen) 2 (qtpy)] 2+ as effective hybridization indicator . Malliaras studied and reviewed transistors (Scheme ) based on PEDOT–PSS for chemo/bio‐sensing applications .…”

Sensitive and Selective Detection of DNA Fragments Associated with Ganoderma Boninense by DNA-Nanoparticle Conjugate Hybridisation

“…Further, their application in the routine analysis is limited by their complex protocols, labile reagents with high cost, relatively low sensitivity, and the requirement of highly skilled personnel. Electrochemical methods for the detection of quinoline (a secondary metabolite produced by the palms under Ganoderma attack) [17], and DNA fragments associated with this organism [18,19] have also been reported. The latter, in particular, has been shown to give high sensitivity (5 mL of analyte at ~ 1 fM of target DNA) and selectivity towards G. boninense.…”

Detection of Quinoline in G. boninense-Infected Plants Using Functionalized Multi-Walled Carbon Nanotubes: A Field Study

“…Some methods have included polymerase chain reaction (PCR) amplification [ 12 , 13 , 14 ] but have encountered challenges of gene bank aberrations resulting from sequencing [ 2 , 13 , 15 ]. Other recent methods have used electrochemical sensors and have involved the complex process of extracting DNA from the infected part of a tree [ 16 , 17 , 18 , 19 ]. The disease has also been detected via hyperspectral data and imagery techniques [ 2 , 20 , 21 ] with the attendant problems of tying to observe palm leaves coupled at the top of palm canopies.…”

“…It bears a positive charge while in solution, thus easily attaching to any substance with a negative charge, and it also displays good film formation, zero toxicity, high mechanical strength, and excellent water permeability [ 27 ]. Gold nanoparticles (AuNPs) are another kind of nanomaterial known for their catalytic ability and particle size-dependent use [ 17 , 19 ]. Modification of carbon nanotubes (CNTs) with CTS and AuNPs has been successfully carried out and reported [ 28 ].…”