Microarrays pp 247-260 | Cite as

eSensor®A Microarray Technology Based on Electrochemical Detection of Nucleic Acids and Its Application to Cystic Fibrosis Carrier Screening

  • Michael R. Reed
  • William A. Coty
Part of the Integrated Analytical Systems book series (ANASYS)


We have developed a test for identification of carriers for cystic fibrosis using the eSensor® DNA detection technology. Oligonucleotide probes are deposited within self-assembled monolayers on gold electrodes arrayed upon printed circuit boards. These probes allow sequence-specific capture of amplicons containing a panel of mutation sites associated with cystic fibrosis. DNA targets are detected and mutations genotyped using a “sandwich” assay methodology employing electrochemical detection of ferrocene-labeled oligonucleotides for discrimination of carrier and non-carrier alleles. Performance of the cystic fibrosis application demonstrates sufficient accuracy and reliability for clinical diagnostic use, and the procedure can be performed by trained medical technologists available in the hospital laboratory.


Cystic Fibrosis Print Circuit Board Signal Probe Capture Probe Cystic Fibrosis Transmembrane Regulator 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



A large number of people have contributed to the development of the eSen-sor® DNA Detection Technology, beginning with the founders of Clinical Micro Sensors, Thomas Meade and Jon Faiz Kayyem, and including past employees of CMS and Motorola Life Sciences as well as current employees of Osmetech Molecular Diagnostics too numerous to mention by name, but whose creativity, enthusiasm, and hard work are gratefully acknowledged.


  1. 1.
    Kerman, K., Kobayashi, M., and Tamiya, E. (2004). Recent trends in electrochemical DNA biosensor technology. Meas. Sci. Technol. 15: R1–R11.CrossRefGoogle Scholar
  2. 2.
    Meade, T.J. and Kayyem, J.F. (1997) Nucleic acid mediated electron transfer. United States Patent 5,591,578.Google Scholar
  3. 3.
    Umek, R.M., Lin, S.W., Vielmetter, J., Terbrueggen, R.H., Irvine, B., Yu, C.J., Kayyem, J.F., Yowanto, H., Blackburn, G.F., Farkas, D.H., and Chen, Y.P. (2001) Electronic detection of nucleic acids: A versatile platform for molecular diagnostics. J. Mol. Diag. 3: 74–84.Google Scholar
  4. 4.
    Yu, C.J., Wan, Y., Yowanto, H., Li, J., Tao, C., James, M.J., Tan, C.L., Blackburn, G.F., and Meade, T.J. (2001) Electronic detection of single-base mismatches in DNA with ferrocene-modified probes. J. Am. Chem. Soc. 123: 11155–11161.CrossRefGoogle Scholar
  5. 5.
    Finklea, H.O. (1998) Electrochemistry of organized monolayers of thiols and related molecules on electrodes. Electroanal. Chem.: Series Adv., 19: 109–335.Google Scholar
  6. 6.
    Estroff, L.A., Kriebel, J.K., Nuzzo, R.G., and Whitesides, G.M. (2005) Self-assembled monolayers of thiolates on metals as a form of nanotechnology. Chem. Rev. 105: 1103–1169.CrossRefGoogle Scholar
  7. 7.
    Eckstein, F. (1991). Oligonucleotides and Analogues: A Practical Approach. IRL Press, New York, 313 pp.Google Scholar
  8. 8.
    Gorton, J.E., Lentzner, H.L., and Watts, W.E. (1971) Bridged ferrocenes-VIII: Polarographic half-wave potentials of ferrocenophanes and related compounds. Tetrahedron, 27: 4353–4360.CrossRefGoogle Scholar
  9. 9.
    Nahir, T.M. and Bowden, E.F.J. (1996) The distribution of standard rate constants for electron transfer between thiol-modified gold electrodes and adsorbed cytochrome c. Electroanal. Chem. 410: 9–13.CrossRefGoogle Scholar
  10. 10.
    Moskowitz, S.M., Gibson, R.L., Sternen, D.L., Cheng, E., Cutting, G.R. (2005) CFTR-Related Disorders. Gene Reviews,
  11. 11.
    Grody, W.W., Cutting, G.R., and Watson, M.S. (2007) The cystic fibrosis mutation “arms race”: When less is more. Genet. Med. 9: 739–744.CrossRefGoogle Scholar
  12. 12.
    Watson, M.S., Cutting, G.R., Desnick, R.J., Driscoll, D.A., Klinger, K., Mennuti, M., Palomaki, G.E., Bradley W. Popovich, B.W., Pratt, V.M., Rohlfs, E.M., Strom, C.M., Richards, C.S., Witt, D.R., and Grody, W.W. (2004) Cystic fibrosis population carrier screening: 2004 revision of American College of Medical Genetics mutation panel. Genet. Med. 6: 387–391.CrossRefGoogle Scholar
  13. 13.
    Grody, W.W., Cutting, G.R., Klinger, K.W., Richards, C.S., Watson, M.S., and Desnick, R.J. (2001) Laboratory standards and guidelines for population-based cystic fibrosis carrier screening. Genet. Med. 3: 149–154.CrossRefGoogle Scholar
  14. 14.
    Box, G.E.P., Hunter, W.G., and Hunter, J.S. (1978) Statistics for Experimenters: An Introduction to Design, Data Analysis and Model Building. John Wiley and Sons, New York, 644 pp.Google Scholar
  15. 15.
    Higuchi, R.G. and Ochman, H. (1989) Production of single-stranded templates by exonuclease digestion following the ploymerase chain reaction. Nucl. Acids Res. 17: 5865.CrossRefGoogle Scholar
  16. 16.
    Amos, J., Feldman, G.L., Grody, W.W., Monaghan, K., Palomaki, G.E., Prior, T.E., Richards, C.S., and Watson, M.S. (2005) Technical Standards and Guidelines for CFTR Mutation Testing,
  17. 17.
    Kilinc, M.O., Ninis, V.N., Tolun, A., Estivill, X., Casals, T., Savov, A., Dagli, E., Karakoc, F., Demirkol, M., Hüner, G., Özkinay, F., Demir, E., Seculi, J.L., Pena, J., Bousono, C., Ferrer-Calvette, J., Calvo, C., Glover, G., and Kremenski, I. (2000) Genotype-phenotype correlation in three homozygotes and nine compound heterozygotes for the cystic fibrosis mutation 2183AA→G shows a severe phenotype. J. Med. Genet., 37: 307–309.CrossRefGoogle Scholar
  18. 18.
    Ferec, C., Audrezet, M.P., Mercier, B., Guillermit, H., Moullier, P., Quere, I., and Verlingue, C. (1992) Detection of over 98% cystic fibrosis mutations in a Celtic population. Nat. Genet. 1: 188–191.CrossRefGoogle Scholar
  19. 19.
    Ferec, C., Novelli, G., Verlingue, C., Quere, I., Dallapiccola, B., Audrezet, M.P., and Mercier, B. (1995) Identification of six novel CFTR mutations in a sample of Italian cystic fibrosis patients. Mol. Cell Probes 9: 135–137.CrossRefGoogle Scholar
  20. 20.
    Delmarco, A., Pradal, U., Cabrini, G., Bonizzato, A., and Mastella, G. (1997) Nasal potential difference in cystic fibrosis patients presenting borderline sweat test. Eur. Respir. J. 10: 1145–1149.CrossRefGoogle Scholar
  21. 21.
    Bisceglia, L., Grifa, A., Zelante, L., and Gasparini, P. (1994) Development of RNA-SSCP protocols for the identification and screening of CFTR mutations: Identification of two new mutations. Hum. Mutat. 4: 136–140.CrossRefGoogle Scholar
  22. 22.
    Schwartz, M., Anvret, M., Claustres, M., Eiken, H.G., Eiklid, K., Schaedel, C., Stolpe, L., and Tranebjaerg, L. (1994) 394delTT: A Nordic cystic fibrosis mutation. Hum. Genet. 93:157–161.CrossRefGoogle Scholar
  23. 23.
    Claustres, M., Laussel, M., Desgeorges, M., Giansily, M., Culard, J.F., Razakatsara, G., and Demaille, J. (1993) Analysis of the 27 exons and flanking regions of the cystic fibrosis gene: 40 different mutations account for 91.2% of the mutant alleles in southern France. Hum. Mol. Genet. 2: 1209–1213.CrossRefGoogle Scholar
  24. 24.
    NCCLS (2002) Interference Testing in Clinical Chemistry; Approved Guideline. NCCLS Document EP7-A. NCCLS, Wayne, PA, 104 pp.Google Scholar
  25. 25.
    Waleed, A.A, Jönsson, L.J., and Rådström, P. (2000) Identification and characterization of immunoglobulin G in blood as a major inhibitor of diagnostic PCR. J. Clin. Micro. 38: 345–350.Google Scholar
  26. 26.
    Waleed, A.A. and Rådström, P. (2001) Purification and characterization of PCR-inhibitory components in blood cells. J. Clin. Micro. 39: 485–493.CrossRefGoogle Scholar
  27. 27.
    Liu, R.H, Yang, J., Lenigk, R. Bonanno, J., and Grodzinski, P. (2004) Self-contained, fully integrated biochip for sample preparation, polymerase chain reaction amplification, and DNA microarray detection. Anal. Chem. 76: 1824–1831.CrossRefGoogle Scholar
  28. 28.
    Liu, R.H., Coty, W. A., Reed, M.R. and Gust, G (2008) Electrochemical detection-based DNA micro arrays. IVD Technology 7: 36–42.Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Michael R. Reed
    • 1
  • William A. Coty
    • 1
  1. 1.Osmetech Molecular DiagnosticsPasadena

Personalised recommendations