Sensitive bioaffinity assays with individual microparticles and time-resolved fluorometry

Clin Chem. 1997 Oct;43(10):1937-43.

Abstract

Future immunoassays and nucleic acid hybridization assays will be performed in miniaturized formats that utilize microchips or microparticles. This will require a sensitive detection technology that allows spatial resolution. By using fluorescent europium chelates and time-resolved microfluorometry, one can detect 11,000 europium molecules on individual microparticles. In a miniaturized noncompetitive immunoassay of prostate-specific antigen (PSA), we quantitatively detected 5 ng/L (0.05 amol per particle) of the analyte on an individual microparticle with excellent precision over the whole measurement range (CV <10%). Using a hybridization assay, we also could detect the deltaF508 mutation for cystic fibrosis on individual microparticles. Consequently, fluorescent lanthanide chelate labels and time-resolved microfluorometry qualify as the next generation of technology in this field.

MeSH terms

  • Chelating Agents
  • Cystic Fibrosis / genetics
  • Europium
  • Humans
  • Immunoassay / standards*
  • Mutation
  • Prostate-Specific Antigen / analysis
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence / methods*
  • Time Factors

Substances

  • Chelating Agents
  • Europium
  • Prostate-Specific Antigen