Base pair mismatch recognition using plasmon resonant particle labels

Anal Biochem. 2002 Oct 1;309(1):109-116. doi: 10.1016/s0003-2697(02)00410-4.

Abstract

We demonstrate the use of silver plasmon resonant particles (PRPs), as reporter labels, in a microarray-based DNA hybridization assay in which we screen for a known polymorphic site in the breast cancer gene BRCA1. PRPs (40-100 nm in diameter) image as diffraction-limited points of colored light in a standard microscope equipped with dark-field illumination, and can be individually identified and discriminated against background scatter. Rather than overall intensity, the number of PRPs counted in a CCD image by a software algorithm serves as the signal in these assays. In a typical PRP hybridization assay, we achieve a detection sensitivity that is approximately 60 x greater than that achieved by using fluorescent labels. We conclude that single particle counting is robust, generally applicable to a wide variety of assay platforms, and can be integrated into low-cost and quantitative detection systems for single nucleotide polymorphism analysis.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Base Pair Mismatch*
  • Base Sequence
  • Biotinylation
  • Breast Neoplasms / genetics
  • DNA / chemistry
  • DNA / genetics*
  • Female
  • Genes, BRCA1
  • Humans
  • Molecular Sequence Data
  • Nucleic Acid Hybridization
  • Oligonucleotide Array Sequence Analysis / methods*
  • Polymorphism, Single Nucleotide
  • Sensitivity and Specificity
  • Surface Plasmon Resonance / instrumentation
  • Surface Plasmon Resonance / methods*
  • Templates, Genetic

Substances

  • DNA