PRINCIPLE AND REVIEW ON DNA BIOSENSOR

Nor Azah Yusof (PhD)
Chemistry Department, Faculty of Science, UPM

OUTLINE
Review on development of different types of DNA based biosensor (Optical/Electrochemical/Magnetic/ Piezo) Advantage and disadvantages of different types of DNA based biosensor

Introduction
Biosensor-small device employing biological recognition properties for a selective bioanalysis. Biosensor-eliminate the need of the sample preparation, hence offer great promise for onsite analytical application.

. antibodies. enzymes or DNA.Introduction The sensing element maybe whole cells.

The current method for the identification of specific DNA sequences are based on isolation of double stranded DNA and further PCR to amplify the target sequence of DNA. .Introduction DNA biosensors are of major interest due to tremendous promise for obtaining sequence specific information in a faster. simpler and cheaper manner compared to the traditional analysis. The target DNA is exposed to labeled probe DNA.

Optical sensors Fluorescence (quantum dot. nanowires. conjugated polymers. graphene) Surface plasma resonance .

Fluorescence Molecular Beacons (MB)– oligonucleotides with stem and loop structure labeled with fluorophore at one end and a quencher on the other end.Optical sensors . MB probes offer high sensitivity and specificity Costly .

limit invivo usage Silicon based QD ease this concern .Optical sensors . which exhibit intriguing electronic transitions that resemble single atom behavior (CdSe. CdS.Fluorescence - Quantum dot (QD) An ultrasensitive nanosensor based on fluorescence resonance energy transfer (FRET) Quantum dots (QDs) are nanocrystals made of semiconductors. ZnO) Toxic.

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Fluorescence Nanowires nanowires functionalized with DNA to specific regions of a photolithographically patterned substrate.Optical sensors . .

The complementary DNA attached to the nanowires shows that the wires assembled in the proper locations.DNA-tagged assembled nanowires have been exposed to complementary DNA that is tagged with fluorescent dyes. .

saliva or another bodily fluid. The researchers would then dip the biosensor hook into treated blood. Like stringing a worm on a hook. scientists would place DNA from a gene that's known to contribute to a disease's development onto a piece of graphene. the biosensor gives off a signal that scientists can detect. If DNA from the disease-causing gene is in the fluid and takes the bait. .Fluorescence A graphene-DNA biosensor would detect diseases by fishing for molecules involved in disease.Optical sensors .

C) A complimentary DNA nears the ssDNA and causes the adsorbed ssDNA to detach from the graphene surface. D) DNA adsorbed onto graphene is protected from being broken down by enzymes.How fluorescent-tagged DNA interacts with functionalized graphene. causing the fluorescence on the ssDNA to darken more. . Both single-stranded DNA (A) and double-stranded DNA (B) are adsorbed onto a graphene surface. but the interaction is stronger with ssDNA.

Biosensors and Bioelectronics.Optical sensors ..polythiophene and polyarylene .Fluorescence Conjugated polymers DNA biosensors based on water-soluble conjugated polymers Liu et al. 26 (2011) 2154– 2164 Derivatives of polyfluorene.

The dry-reagent strip type biosensor has been developed for visual detection of double stranded DNA within a short time. .Colorimetric or Strip type DNA Sensor A novel nanoparticle based colorimetric detection offers great promise for direct detection of DNA hybridization. In this case. results in changes of the optical properties of the aggregated functional gold nanoparticles. Oligo nucleotide conjugated gold nanoparticle is used as probe for detection of target DNA through hybridization. a distance change. occurred from the hybridization event.

Colorimetric or Strip type DNA Sensor .

Optical sensors . .SPR Surface Plasmon Resonance .Quantum optical electrical phenomenon arising from the interaction of light with metal surface. - Change in resonance angle due to change in the interfacial refractive index resulting from the surface binding reaction. Energy transfer occurs only at specific resonance wavelength of light.

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Amperometric The hybridization is commonly detected by the increase in current signal due to redox indicator (organic dyes. Inc.Electrochemical DNA Biosensors (Voltammetry. metal complexes or enzymes) that recognizes the DNA duplex. Redox-active molecules based two commercialized DNA chips have been introduced in molecular diagnosis market in the trade name of eSensorTM produced by Motorola Life sciences . and GenlyserTM by Toshiba. DPV. Conductometric. . Dielectric) CV.

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Electrochemical DNA Biosensors – sensitized by nanoparticles Ultrasensitive Electrochemical Detection of DNA Based on PbS Nanoparticle Tags and Nanoporous Gold Electrode Electrochemical DNA biosensor for the detection of DNA hybridization with the amplification of Au nanoparticles and CdS nanoparticles .

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(d) bare modified electrode .DPV curves obtained in Tris–HCl aqueous solution using Co(phen)2 2+ as an electrochemical hybridization label for detection of different DNA sequences.018×10−10 M complementary ssDNA. . (a)6. (b)6.018×10−8 M one mismatched ssDNA. (c) capture ssDNA.

(d) 8.006×10−10 M.The DPV signals obtained for different concentrations of target DNA: The concentration of target DNA: (a) 6. . (f) 0.018×10−11 M. (c) 2.024×10−11 M. (e) 6. (b) 4.012×10−10 M.018×10−10 M.

poly(vinylferrocenium) perchlorate (PVF+ClO4-) coated as a film on Pt electrodes.Electrochemical DNA Biosensors – sensitized by conducting polymer Electrochemical detection of DNA hybridization based on polypyrrole/ssDNA/multi-wall carbon nanotubes paste electrode Characterization of redox polymer. and the detection of DNA based on the electrochemical behavior of the polymer .

A nanogap dielectric biosensor .

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The low frequency response indicates the largescale molecular structure changes like the conformation changes of DNA during hybridization from single stranded DNA (ss-DNA) to double stranded DNA. they respond differently depending on the frequency. . and the largescale molecular structure.The dielectric properties of molecules depend on electron transfer. When an oscillatory field perturbs biomolecules. atomic bonds.

causing an increase in conductance . Target molecules can be recognized by the receptor modified on the channel surface through strong binding affinity.Conductometric (a) The illustration of a nanoscale FET biosensor with a cross-sectional view. negatively charged target molecules captured by the receptor would make an accumulation of hole carriers. (b) When positively charged target molecules bind the receptor modified on a p-type NW. The semiconductor channel (NW or NT) is placed between the source and drain electrodes with a gate electrode on the bottom to modulate the conductivity of the semiconductor channel. On the contrary. resulting in a decrease in conductance. positive carriers (holes) are depleted in the NW.

(d) CgA was released from neurons stimulated by glutamate and was detected by CgA-Ab/CNT-FET. Immediately after the glutamate (50 M) stimulation.(c) Schematic representation of a CNT-FET device including the surface modification and molecular recognition procedures: (1) modification of linkers onto the single-walled CNT through — interaction. a dramatic increase in current was detected due to the binding of the released CgA to CgAAb/CNTFET . (2) immobilization of antibody. A coverslip with grown neurons was positioned on the CgA-Ab/CNT-FET device with neurons facing the FET circuits. (3) detection of antigen by antibody.

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Issue 2. CHINESE JOURNAL OF ANALYTICAL CHEMISTRY.MAGNETIC BEAD-BASED DNA DETECTION Via electrochemical detector Fang et al. Volume 37.. February 2009 .

Via Fluorescence .

Piezoelectric DNA biosensor .

cost. . associated with the hybridization reaction. results in a decrease of the oscillating frequency. The quartz crystal microbalance (QCM) is an extremely sensitive massmeasuring device that allows dynamic monitoring of hybridization events. sensitivity and real time label-free detection.Piezoelectric DNA biosensor is based on quartz crystal that oscillate at a defined frequency when an oscillating voltage is applied. QCM hybridization biosensors consist of an oscillating crystal with the DNA probe immobilized on its surface. The increased mass. allowing high sensitivity. Piezoelectric method has recently emerged as most attractive due to their simplicity.

amplification.Conclusions and Future Prospects From the first discovery of electrochemistry of nucleic acids by Palecek at the end of the 1950’s [81]. and conversion of the biological signals . processing. Future biosensors will require the development of new reliable devices or the improvement of the existing ones in order to allow superior transduction. particularly at the development of electrochemical DNA biosensors based on the nucleic acid as biorecognition element The use of DNA biostrip and biochip technologies eliminates the role of PCR.huge progress can be observed.

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