Encoded metal nanoparticle-based molecular beacons for multiplexed detection of DNA

Michael Y. Sha, Mark Yamanaka, Ian D. Walton, Scott M. Norton, Rebecca L. Stoermer, Christine D. Keating, Michael J. Natan, Sharron G. Penn

Research output: Contribution to journalArticle

21 Scopus citations

Abstract

In this paper we describe a molecular beacon format assay in which encoded nanowire particles are used to achieve multiplexing. We demonstrate this principle with the detection of five viral pathogens; Hepatitis A virus, Hepatitis C virus, West Nile Virus, Human Immune Deficiency virus and Severe Acute Respiratory Syndrome virus. Oligonucleotides are designed complementary to a target sequence of interest containing a 3′ universal fluorescence dye. A 5′ thiol causes the oligonucleotides to self-assemble onto the metal nanowire. The single-stranded oligonucleotide contains a self-complementary hairpin stem sequence of 10 bases that forces the 3′ fluorophore to come into contact with the metallic nanowire surface, thereby quenching the fluorescence. Upon addition of target DNA, there is hybridization with the complementary oligonucleotides. The resulting DNA hybrid is rigid, unfolds the hairpin structure, and causes the fluorophore to be moved away from the surface such that it is no longer quenched. By using differently encoded nanowires, each conjugated with a different oligonucleotide sequence, multiplexed DNA assays are possible using a single fluorophore, from a multiplexed RT-PCR reaction.

Original languageEnglish (US)
Pages (from-to)327-335
Number of pages9
JournalNanobiotechnology
Volume1
Issue number4
DOIs
StatePublished - 2005

All Science Journal Classification (ASJC) codes

  • Bioengineering
  • Biomedical Engineering
  • Molecular Biology

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    Sha, M. Y., Yamanaka, M., Walton, I. D., Norton, S. M., Stoermer, R. L., Keating, C. D., Natan, M. J., & Penn, S. G. (2005). Encoded metal nanoparticle-based molecular beacons for multiplexed detection of DNA. Nanobiotechnology, 1(4), 327-335. https://doi.org/10.1385/NBT:1:4:327