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