Comparison of fluorescence and resonance light scattering for highly sensitive microarray detection of bacterial pathogens

J Microbiol Methods. 2003 Dec;55(3):755-62. doi: 10.1016/j.mimet.2003.08.008.

Abstract

Microarrays have emerged as potential tools for bacterial detection and identification. Given their high parallelism, they might represent a breakthrough in current diagnostic methods, provided they can be coupled to simplified labeling protocols and detected with adequate sensitivities. We describe here a technique to directly label total bacterial RNA, thus avoiding the multiple steps and possible biases associated with enzymatic amplification (e.g. PCR). We have then compared the performances of one white-light source and two laser-based fluorescence scanners for detection reliability and sensitivity. Our study reveals that nanoparticle-labeled bacterial RNA generates reproducible resonance light scattering signals that are at least 50 times more intense than state-of-the-art confocal-based fluorescence signals.

Publication types

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

MeSH terms

  • Light
  • Microscopy, Fluorescence
  • Oligonucleotide Array Sequence Analysis / methods*
  • RNA, Bacterial / chemistry
  • RNA, Bacterial / genetics
  • Reproducibility of Results
  • Scattering, Radiation*
  • Sensitivity and Specificity
  • Staphylococcus aureus / genetics*
  • Staphylococcus aureus / isolation & purification

Substances

  • RNA, Bacterial