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Electrically detected displacement assay (EDDA): a practical approach to nucleic acid testing in clinical or medical diagnosis

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Abstract

This paper introduces the electrically detected displacement assay (EDDA), a electrical biosensor detection principle for applications in medical and clinical diagnosis, and compares the method to currently available microarray technologies in this field. The sensor can be integrated into automated systems of routine diagnosis, but may also be used as a sensor that is directly applied to the polymerase chain reaction (PCR) reaction vessel to detect unlabeled target amplicons within a few minutes. Major aspects of sensor assembly like immobilization procedure, accessibility of the capture probes, and prevention from nonspecific target adsorption, that are a prerequisite for a robust and reliable performance of the sensor, are demonstrated. Additionally, exemplary results from a human papillomavirus assay are presented.

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Abbreviations

ACV:

alternating current voltammetry

CP:

capture probe

CPG:

controlled pore glass

CV:

cyclic voltammetry

DBU:

diazabicycloundecene

DMAP:

4-dimethylaminopyridine

DMF:

dimethylformamide

DMSO:

dimethylsulfoxide

ds:

double stranded

DTE:

dithioerythritol

DTPA:

dithiolphosphoamidite

EC:

electrochemical

EDDA:

electrically detected displacement assay

ET:

electron transfer

FeAc:

ferroceneacetic acid

HATU:

O-azabenzotriazol-1-yl-tetramethyluronium hexafluorophosphate

HBTU:

O-benzotriazol-1-yl-tetramethyluronium hexafluorophosphate

HPV:

human papillomavirus

LCAA-CPG:

long-chain alkylamine controlled pore glass

MEA:

microelectrode array

MMTr:

monomethoxytrityl

NHS:

N-hydroxysuccinimide

pcb:

printed circuit board

PCR:

polymerase chain reaction

PEEK:

polyetheretherketone

rCP:

reference capture probe

rSP:

reference signalling probe

SP:

signalling probe

ss:

single stranded

T:

target

TEAA:

triethyl ammonium acetate

THF:

tetrahydrofuran

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Acknowledgements

Part of this work was financially supported by a governmental grant (sponsored by bmbf, program BioChance plus, grant 0313611A) which is gratefully acknowledged.

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Authors and Affiliations

  1. FRIZ Biochem Gesellschaft für Bioanalytik mbH, 82061, Neuried, Germany

    P. Liepold, T. Kratzmüller, N. Persike, M. Bandilla, M. Hinz, H. Wieder, H. Hillebrandt, E. Ferrer & G. Hartwich

  2. Roche Diagnostics GmbH, Sandhofer Str. 116, 68305, Mannheim, Germany

    H. Wieder

  3. Roche Diagnostics GmbH, Nonnenwwlad 2, 82377, Penzberg, Germany

    H. Hillebrandt

  4. Kendle GmbH + Co.GMI KG, Stefan-George-Ring 6, 81929, München, Germany

    E. Ferrer

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  1. P. Liepold
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  2. T. Kratzmüller
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  4. M. Bandilla
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  9. G. Hartwich
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Correspondence to G. Hartwich.

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Liepold, P., Kratzmüller, T., Persike, N. et al. Electrically detected displacement assay (EDDA): a practical approach to nucleic acid testing in clinical or medical diagnosis. Anal Bioanal Chem 391, 1759–1772 (2008). https://doi.org/10.1007/s00216-008-2045-5

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  • DOI: https://doi.org/10.1007/s00216-008-2045-5

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