Bienzymatic-based electrochemical DNA biosensors: a way to lower the detection limit of hybridization assays
Literature Information
Murielle Rochelet-Dequaire, Naïma Djellouli, Benoît Limoges, Pierre Brossier
The use of the alkaline phosphatase (AP) as an enzyme label and the amplification of its analytical response with a diaphorase (DI) secondary enzyme were investigated in an electrochemical hybridization assay involving arrays of carbon screen-printed DNA biosensors for the sensitive quantification of an amplified 406-base pair human cytomegalovirus DNA sequence (HCMV DNA). For this purpose, PCR-amplified biotinylated HCMV DNA targets were simultaneously bound to a monolayer of neutravidin irreversibly adsorbed on the surface of the electrodes and hybridized to complementary digoxigenin-labeled detection probes. The amount of hybrids immobilized on the electrode surface was labeled with an anti-digoxigenin AP conjugate and quantified electrochemically by measuring the activity of the AP label through the hydrolysis of the electroinactive p-aminophenylphosphate (PAPP) substrate into the p-aminophenol (PAP) product. The intensity of the cyclic voltammetric anodic peak current resulting from the oxidation of PAP into p-quinoneimine (PQI) was related to the number of viral amplified DNA targets present in the sample, and a detection limit of 10 pM was thus achieved. The electrochemical response of the AP label product was further enhanced by adding the diaphorase enzymatic amplifier in the solution. In the presence of the auxiliary enzyme DI, the PQI was reduced back to PAP and the resulting oxidized form of DI was finally regenerated in its reduced native state by its natural substrate, NADH. Such a bienzymatic amplification scheme enabled a 100-fold lowering of the HCMV DNA detection limit obtained with the monoenzymatic system.
Recommended Journals

Acta Materialia

Current Opinion in Colloid & Interface Science

Russian Journal of General Chemistry

Journal of Peptide Science

Chemistry Education Research and Practice

Russian Journal of Organic Chemistry

Russian Journal of Coordination Chemistry

Journal of Natural Medicines

Current Opinion in Solid State & Materials Science

Crystallography Reports
Related Literature
Organic nanoparticles of malachite green with enhanced far-red emission: size-dependence of particle rigidity
Tomohito Funada, Takuya Hirose, Naoto Tamai, Hiroshi Yao
DOI: 10.1039/C5CP00031A
Molecular thermodynamics of metabolism: quantum thermochemical calculations for key metabolites
C. Panayiotou
DOI: 10.1039/C4CP05825A
Enhancing electrochemical performance by control of transport properties in buffer layers – solid oxide fuel/electrolyser cells
Devaraj Ramasamy, Narendar Nasani, Ana D. Brandão, Domingo Pérez Coll, Duncan P. Fagg
DOI: 10.1039/C5CP00778J
Atomic collisions in suprafluid helium-nanodroplets: timescales for metal-cluster formation derived from He-density functional theory
Andreas W. Hauser, Alexander Volk, Philipp Thaler, Wolfgang E. Ernst
DOI: 10.1039/C5CP01110H
Comment on “How to interpret Onsager cross terms in mixed ionic electronic conductors” by I. Riess, Phys. Chem. Chem. Phys., 2014, 16, 22513
Han-Ill Yoo, Manfred Martin, Juergen Janek
DOI: 10.1039/C4CP05737F
Order and disorder around Cr3+ in chromium doped persistent luminescent AB2O4 spinels
Neelima Basavaraju, Kaustubh R. Priolkar, Didier Gourier, Aurélie Bessière, Bruno Viana
DOI: 10.1039/C5CP01097G
Vibrational models for a crystal with 36 water molecules in the unit cell: IR spectra from experiment and calculation
Pavlin D. Mitev, Anders Eriksson, Jean-François Boily, Kersti Hermansson
DOI: 10.1039/C5CP00390C
Enhancement of the thermoelectric figure of merit in DNA-like systems induced by Fano and Dicke effects
Hua-Hua Fu, Lei Gu, Dan-Dan Wu, Zu-Quan Zhang
DOI: 10.1039/C4CP04382K
Fabrication of charged membranes by the solvent-assisted lipid bilayer (SALB) formation method on SiO2 and Al2O3
DOI: 10.1039/C5CP01428J
Towards a comprehensive insight into efficient hydrogen production by self-assembled Ru(bpy)32+–polymer–Pt artificial photosystems
Huan Lin, Dan Liu, Jinlin Long, Zizhong Zhang, Huaqiang Zhuang, Yi Zheng, Xuxu Wang
DOI: 10.1039/C5CP00720H
You might also like
Is 2-(2-chloroacetamido)-3-phenylpropanoic acid (CAS: 7765-11-9) safe?
2-(2-Chloroacetamido)-3-phenylpropanoic acid (CAS: 7765-11-9) is generally consi...
Is 2-(Benzyloxy)-5-bromobenzoic acid (CAS: 62176-31-2) safe?
2-(Benzyloxy)-5-bromobenzoic acid can be handled safely if appropriate precautio...
What is (4-Methyl-1,2,5-oxadiazol-3-yl)methanamine hydrochloride (CAS: 1159825-48-5)?
(4-Methyl-1,2,5-oxadiazol-3-yl)methanamine hydrochloride is a chemical compound ...
What is 2-(5-Hexylthiophen-2-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (CAS: 917985-54-7)?
2-(5-Hexylthiophen-2-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (CAS: 917985-54...
Are there alternatives to 4-(8-Methyl-9H-1,3-dioxolo[4,5-h][2,3]benzodiazepin-5-yl)benzenamine (CAS: 102771-26-6) in synthesis?
While 4-(8-Methyl-9H-1,3-dioxolo[4,5-h][2,3]benzodiazepin-5-yl)benzenamine (CAS:...
What is the market or research trend for tert-butyl 3-hydroxy-4,5,7,8-tetrahydro-2H-pyrazolo[3,4-d]azepine-6-carboxylate (CAS: 851376-80-2)?
The market for tert-butyl 3-hydroxy-4,5,7,8-tetrahydro-2H-pyrazolo[3,4-d]azepine...
How should waste containing 3,5-Diamino-1H-pyrazole-4-carbonitrile (CAS: 6844-58-2) be handled?
Waste containing 3,5-Diamino-1H-pyrazole-4-carbonitrile (CAS: 6844-58-2) should ...
How is (6-Fluoro-3-pyridinyl)boronic acid (CAS: 351019-18-6) typically synthesized?
(6-Fluoro-3-pyridinyl)boronic acid can be synthesized through the reaction of 6-...
What industries use Dibenzyl carbonimidoylbiscarbamate (CAS: 10065-79-9)?
Dibenzyl carbonimidoylbiscarbamate (CAS: 10065-79-9) finds applications in vario...
What is the market or research trend for (beta,beta,2,3,4,5,6-~2~H_7_)Phenylalanine (CAS: 74228-83-4)?
The market for (beta,beta,2,3,4,5,6-~2~H_7_)Phenylalanine (CAS: 74228-83-4) is g...
Source Journal
Analyst

Analyst publishes analytical and bioanalytical research that reports premier fundamental discoveries and inventions, and the applications of those discoveries, unconfined by traditional discipline barriers.




![3-[4-(difluoromethoxy)phenyl]-2-({[(9H-fluoren-9-yl)methoxy]carbonyl}amino)propanoic acid structure 3-[4-(difluoromethoxy)phenyl]-2-({[(9H-fluoren-9-yl)methoxy]carbonyl}amino)propanoic acid structure](https://static.chemtradehub.com/structs/149/1496564-27-2-952e.webp)