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Clinical Chemistry 45: 1659-1664, 1999;
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(Clinical Chemistry. 1999;45:1659-1664.)
© 1999 American Association for Clinical Chemistry, Inc.

Articles

Thin Film Biosensor for Rapid Visual Detection of Nucleic Acid Targets

Rachel M. Ostroffa, Deborah Hopkins, Ayla B. Haeberli, Wahab Baouchi and Barry Polisky

BioStar, Inc., 6655 Lookout Rd., Boulder, CO 80301.
a Author for correspondence. Fax 303-530-6627; e-mail rmo{at}csn.net

Background: We have developed a silicon-based biosensor that generates a visual signal in response to nucleic acid targets.

Methods: In this system, capture oligonucleotide probes are immobilized on the surface of the biosensor. Interaction of the capture probes with a complementary target and a biotinylated detector oligonucleotide allows initiation of formation of an organic thin film on the biosensor. Thin film formation is completed by enzymatic activity of peroxidase conjugated to an anti-biotin antibody. Peroxidase catalyzes deposition of an insoluble product onto the silicon surface, generating a uniform thin film. The increased thickness on the surface alters the perceived color of the biosensor through changes in the interference patterns of reflected light from the surface, causing a color change from gold to purple.

Results: The biosensor results may be evaluated by direct visual inspection or quantified by ellipsometry. Results are obtained in 25 min with a detection limit of 5 pmol/L (150 amol/sample). Selectivity of the biosensor is demonstrated by discrimination of single nucleotide mismatches. Multitarget arrays are also analyzed with the thin film biosensor, and the system is capable of detecting targets from human serum and urine.

Conclusions: The biosensor surface is inexpensive to produce, and the assay format is simple and rapid. The thin film biosensor is adaptable to a wide variety of nucleic acid detection applications, including rapid diagnostic testing for infectious disease panels, antibiotic resistance panels, or allelic discrimination of specific genetic markers.




The following articles in journals at HighWire Press have cited this article:


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H. Li and L. Rothberg
Colorimetric detection of DNA sequences based on electrostatic interactions with unmodified gold nanoparticles
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F. Fixe, V. Chu, D. M. F. Prazeres, and J. P. Conde
An on-chip thin film photodetector for the quantification of DNA probes and targets in microarrays
Nucleic Acids Res., May 17, 2004; 32(9): e70 - e70.
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R. Jenison, H. La, A. Haeberli, R. Ostroff, and B. Polisky
Silicon-based Biosensors for Rapid Detection of Protein or Nucleic Acid Targets
Clin. Chem., October 1, 2001; 47(10): 1894 - 1900.
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R. Jenison, A. Haeberli, S. Yang, B. Polisky, and R. Ostroff
Thin Film Biosensor for Rapid Detection of mecA from Methicillin-resistant Staphylococcus aureus
Clin. Chem., September 1, 2000; 46(9): 1501 - 1504.
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Copyright © 1999 by the American Association for Clinical Chemistry.