Applied Microbiology and Biotechnology

, Volume 77, Issue 5, pp 1181–1188 | Cite as

Electrical DNA-chip-based identification of different species of the genus Kitasatospora

  • Robert Möller
  • Thomas Schüler
  • Sebastian Günther
  • Marc René Carlsohn
  • Thomas Munder
  • Wolfgang Fritzsche


The identification of different Kitasatospora strains has been shown with a DNA-chip based on an electrical readout scheme. The 16S-23S rDNA internal transcribed spacer region of these Actinomycetes was used for identification. Two different capture probes per strain were immobilized on the chip. The capture probes were spotted on a DNA-chip with electrode structures for an electrical DNA detection. A biotinylated PCR product of the 16S-23S rDNA region was incubated on the chips and bound to its complementary capture sequences. Followed by a gold nanoparticle or enzyme labeling and a deposition of silver, the binding of the PCR product was detected by an increase of the measured conductivity on the chip. To show the applicability of this detection system, four strains of Kitasatospora were chosen for an identification using the DNA-chip with electrical detection. Each strain was clearly identified using the system. Concentrations of the polymerase chain reaction (PCR) products within the range of 1 ng/ml to 1 μg/ml were detected and identified. These tests are the first application of this novel electrical detection scheme for the identification and classification of microorganisms. The presented results show that the DNA-chip with electrical detection can be used for a robust and cost-efficient DNA analysis.


DNA-chip Electrical readout Kitasatospora Gold nanoparticles 16S-23S rDNA 



We would like to thank R.D. Powell and J.F. Hainfeld from Nanoprobes (Yaphank, NY, USA) for their constant help in establishing the enzymatic silver deposition on the DNA-chips with electrical detection and for providing the EnzMet™ reagents supported by NIH SBIR grant 2R44 GM064257-02. Funding by the DFG (Fr 1348/5–2) is acknowledged.


  1. Cheung VG, Morley M, Aguilar F, Massimi A, Kucherlapati R, Childs G (1999) Making and reading microarrays. Nat Genet 21(1 Suppl):15–19CrossRefGoogle Scholar
  2. Drummond TG, Hill MG, Barton JK (2003) Electrochemical DNA sensors. Nat Biotechnol 21(10):1192–1199CrossRefGoogle Scholar
  3. Fare TL, Coffey EM, Dai H, He YD, Kessler DA, Kilian KA, Koch JE, LeProust E, Marton MJ, Meyer MR, Stoughton RB, Tokiwa GY, Wang Y (2003) Effects of atmospheric ozone on microarray data quality. Anal Chem 75(17):4672–4675CrossRefGoogle Scholar
  4. Fritzsche W, Taton TA (2003) Metal Nanoparticles as Labels for Heterogeneous, Chip-Based DNA Detection. Nanotechnology 14:R63–R73CrossRefGoogle Scholar
  5. Günther S, Groth I, Grabley S, Munder T (2006) Design and evaluation of an oligonucleotide-microarray for the detection of different species of the genus Kitasatospora. J Microbiol Methods 65:226–236CrossRefGoogle Scholar
  6. Gurtler V, Stanisich VA (1996) New approaches to typing and identification of bacteria using the 16S-23S rDNA spacer region. Microbiology 142(Pt 1):3–16CrossRefGoogle Scholar
  7. Hacker GW, Graf AH, Hauser-Kronberger C, Wirnsberger G, Schiechl A, Bernatzky G, Wittauer U, Su HC, Adam H, Thurner J et al (1993) Application of silver acetate autometallography and gold-silver staining methods for in situ DNA hybridization. Chin Med J (Engl) 106(2):83–92Google Scholar
  8. He L, Musick MD, Nicewarner SR, Salinas FG, Nekovic SJ, Natan MJ, Keating CD (2000) Colloidal Au-enhanced surface plasmon resonance for ultrasensitive detection of DNA hybridization. J Am Chem Soc 122:9071–9077CrossRefGoogle Scholar
  9. Li J, Xue M, Lu Z, Zhang Z, Feng L, Chan M (2003) A high-density conduction-based micro-DNA identification array fabricated with a CMOS compatible process. IEEE Trans Electron Devices 50(10):2165–2170CrossRefGoogle Scholar
  10. Möller R, Csaki A, Köhler JM, Fritzsche W (2000) DNA probes on chip surfaces studied by scanning force microscopy using specific binding of colloidal gold. Nucleic Acids Res 28:e91CrossRefGoogle Scholar
  11. Möller R, Csaki A, Köhler JM, Fritzsche W (2001) Electrical classification of the concentration of bioconjugated metal colloids after surface adsorption and silver enhancement. Langmuir 17:5426–5430CrossRefGoogle Scholar
  12. Möller R, Powell RD, Hainfeld JF, Fritzsche W (2005) Enzymatic control of metal deposition as key step for a low-background electrical detection for DNA chips. Nano Lett 5(7):1475–1482CrossRefGoogle Scholar
  13. Ochi K, Hiranuma H (1994) A taxonomic review of the genera Kitasatosporia and Streptoverticillium by analysis of ribosomal protein AT-L30. Int J Syst Bacteriol 44(2):285–292Google Scholar
  14. Omura S, Takahashi Y, Iwai Y, Tanaka H (1982) Kitasatosporia, a new genus of the order Actinomycetales. J Antibiot (Tokyo) 35(8):1013–1019Google Scholar
  15. Park SJ, Taton TA, Mirkin CA (2002) Array-based electrical detection of DNA with nanoparticle probes. Science 295(5559):1503–1506Google Scholar
  16. Prauser H, Schütze B, Martin K (1987) IMET (National collection of microorganisms) catalogue of strains ZIMET, JenaGoogle Scholar
  17. Schaupp CJ, Jiang G, Myers TG, Wilson MA (2005) Active mixing during hybridization improves the accuracy and reproducibility of microarray results. Biotechniques 38(1):117–119Google Scholar
  18. Schena M, Shalon D, Heller R, Chai A, Brown PO, Davis RW (1996) Parallel human genome analysis: Microarray-based expression monitoring of 1000 genes. Proc Natl Acad Sci U S A 93:10614–10619CrossRefGoogle Scholar
  19. Storhoff JJ, Elghanian R, Mucic RC, Mirkin CA, Letsinger RL (1998) One-pot colorimetric differentiation of polynucleotides with single base imperfections using gold nanoparticle probes. J Am Chem Soc 120:1959–1964CrossRefGoogle Scholar
  20. Urban M, Möller R, Fritzsche W (2003) A paralleled readout system for an electrical DNA-hybridization assay based on a microstructured electrode array. Rev Sci Instr 74:1077–1081CrossRefGoogle Scholar
  21. Wellington EM, Stackebrandt E, Sanders D, Wolstrup J, Jorgensen NO (1992) Taxonomic status of Kitasatosporia, and proposed unification with Streptomyces on the basis of phenotypic and 16S rRNA analysis and emendation of Streptomyces Waksman and Henrici 1943, 339AL. Int J Syst Bacteriol 42(1):156–160Google Scholar
  22. Zhang Z, Wang Y, Ruan J (1997) A proposal to revive the genus Kitasatospora (Omura, Takahashi, Iwai, and Tanaka 1982). Int J Syst Bacteriol 47(4):1048–1054CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Robert Möller
    • 3
  • Thomas Schüler
    • 3
  • Sebastian Günther
    • 2
    • 4
  • Marc René Carlsohn
    • 2
  • Thomas Munder
    • 2
    • 5
  • Wolfgang Fritzsche
    • 1
  1. 1.Institute of Photonic TechnologyJenaGermany
  2. 2.Leibniz-Institute for Natural Product Research and Infection Biology e.V.–Hans-Knöll-InstituteJenaGermany
  3. 3.Institute for Physical ChemistryFriedrich-Schiller-UniversityJenaGermany
  4. 4.Department of Veterinary MedicineFreie Universität BerlinBerlinGermany
  5. 5.CLONDIAG Chip TechnologiesJenaGermany

Personalised recommendations