Sensitivity, specificity, and the hybridization isotherms of DNA chips

Biophys J. 2004 Feb;86(2):718-30. doi: 10.1016/S0006-3495(04)74150-8.

Abstract

Competitive hybridization, at the surface and in the bulk, lowers the sensitivity of DNA chips. Competitive surface hybridization occurs when different targets can hybridize with the same probe. Competitive bulk hybridization takes place when the targets can hybridize with free complementary chains in the solution. The effects of competitive hybridization on the thermodynamically attainable performance of DNA chips are quantified in terms of the hybridization isotherms of the spots. These relate the equilibrium degree of the hybridization to the bulk composition. The hybridization isotherm emerges as a Langmuir isotherm modified for electrostatic interactions within the probe layer. The sensitivity of the assay in equilibrium is directly related to the slope of the isotherm. A simpler description is possible, in terms of c(50) values specifying the bulk composition corresponding to 50% hybridization at the surface. The effects of competitive hybridization are important for the quantitative analysis of DNA chip results, especially when used to study point mutations.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Binding Sites
  • Computer Simulation
  • DNA / chemistry*
  • DNA Probes / chemistry*
  • Equipment Failure Analysis / methods*
  • Models, Chemical*
  • Molecular Sequence Data
  • Nucleic Acid Conformation
  • Nucleic Acid Hybridization / methods*
  • Oligonucleotide Array Sequence Analysis / instrumentation*
  • Oligonucleotide Array Sequence Analysis / methods*
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Temperature

Substances

  • DNA Probes
  • DNA