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Impedimetric Detection of DNA Hybridization: Towards Near-Patient DNA Diagnostics

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Immobilisation of DNA on Chips I

Part of the book series: Topics in Current Chemistry ((TOPCURRCHEM,volume 260))

Abstract

Immobilization strategies for the attachment of nucleotide probes to both microarrays and microfabricated interdigitated electrodes differ to address the specific requirements for optical and electrochemical detection, respectively. The DNA immobilization chemistry dictates how the probe molecule is presented to its complement during hybridization and thereby contributes significantly to the final detection signal. This chapter introduces the relevant immobilization strategies for DNA probes on both microarrays and microfabricated interdigitated microsensor electrode arrays. Specifically, we examine immobilization via electrostatic attraction and covalent coupling. The immobilization of DNA presents many challenges because of the need to promote efficient hybridization while minimizing or eliminating nonspecific adsorption to exposed areas of the device substrate. The immobilization strategies also present challenges because of the various materials and surface chemistries that may be involved, the flow characteristics during detection, and the need to perform hybridizations under specific buffer and temperature cycling conditions. This paper provides a rationale for the move towards low-density genosensors using electrochemical detection with focused example applications in human health care. Specifically, electrochemical impedance spectroscopy (EIS) is introduced and discussed as an advantageous method for DNA hybridization detection. The various immobilization strategies are discussed in reference to EIS detection. In conclusion, the advantages of impedance detection are addressed with critical assessments for the advancement of impedance detection for DNA hybridization.

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Abbreviations

A:

adenine

Ag/AgCl:

silver/silver chloride

AC:

alternating current

AFM:

atomic force microscopy

APS:

3-aminopropyltrimethoxysilane

BSA:

bovine serum albumin

C:

cytosine

C :

capacitance

CEPs:

conducting electroactive polymers

CLIA:

Clinical Laboratory Improvement Amendments

CE:

counter electrode

Cy3:

cyanine 3

Cy5:

cyanine 5

DSG:

disuccinimidylglutarate

DNA:

deoxyribose nucleic acid

dsDNA:

double-strand DNA

EIS:

electrical impedance spectroscopy

ATR-FTIR:

attenuated total reflectance-Fourier transform infrared spectroscopy

G:

guanine

GPS:

glycidoxypropyltrimethoxysilane

i :

AC current response signal

i m :

Maximum AC current response signal

IME:

interdigitated microsensor electrode

MPS:

3-mercaptopropyltrimethoxysilane

PAMAM:

polyamidoamine

PDITC:

phenylenediisothiocyanate

PDITC:

phenylenediisothiocyanate

p-t-p:

peak-to-peak

R :

resistance

RE:

reference electrode

RNA:

ribonucleic acid

R o :

equivalent circuit resistance

R s :

solution resistance

SA:

succinic anhydride

ssDNA:

single-strand DNA

SMPB:

succinimidyl 4-[malemidophenyl] butyrate

t :

time

T:

thymine

QCM:

quartz crystal microbalance

V :

voltage

V m :

maximum voltage

WE:

working electrode

X :

reactance

Z :

impedance

Z′:

imaginary component of impedance

Z′′:

real component of impedance

∣Z∣:

magnitude of impedance

θ:

phase shift

τp :

time constant

ω:

radial frequency

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Acknowledgments

This work was supported by the Commonwealth of Virginia's Department of Planning Commonwealth Technologies Research Fund (CTRF grant no. SE2002-02 for Cancer Genomics and Development of Diagnostic Tools and Therapies) and the consortium of the VCU Center for Bioelectronics, Biosensors and Biochips.

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Authors and Affiliations

  1. Center for Bioelectronics, Biosensors, and Biochips (C3B), Department of Chemical and Life Science Engineering, Virginia Commonwealth University, 601 West Main Street, 23284-3038, Richmond, VA, USA

    Anthony Guiseppi-Elie & Louise Lingerfelt

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  1. Anthony Guiseppi-Elie
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  2. Louise Lingerfelt
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Correspondence to Anthony Guiseppi-Elie .

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Christine Wittmann

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Guiseppi-Elie, A., Lingerfelt, L. Impedimetric Detection of DNA Hybridization: Towards Near-Patient DNA Diagnostics. In: Wittmann, C. (eds) Immobilisation of DNA on Chips I. Topics in Current Chemistry, vol 260. Springer, Berlin, Heidelberg. https://doi.org/10.1007/128_006

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