Application of fluorescent nanocrystals (q-dots) for the detection of pathogenic bacteria by flow-cytometry

J Fluoresc. 2010 Jan;20(1):389-99. doi: 10.1007/s10895-009-0546-z. Epub 2009 Oct 14.

Abstract

Fluorescent semiconductor nanocrystals (q-dots) benefit from practical features such as high fluorescence intensity, broad excitation band and emission diameter dependency. These unique spectroscopic characterizations make q-dots excellent candidates for new fluorescent labels in multi-chromatic analysis, such as Flow-Cytometry (FCM). In this work we shall present new possibilities of multi-labeling and multiplex analysis of pathogenic bacteria, by Flow-Cytometry (FCM) analysis and new specific IgG-q-dots conjugates. We have prepared specific conjugates against B. anthracis spores (q-dots585-IgGalphaB. anthracis and q-dots655-IgGalphaB.anthracis). These conjugates enabled us to achieve double staining of B. anthracis spores which improve the FCM analysis specificity versus control Bacillus spores. Moreover, multiplexed analysis of B. anthracis spores and Y. pestis bacteria was achieved by using specific antibodies labeled with different q-dots to obtain: q-dots585-IgGalphaB. anthracis and q-dots655-IgGalphaY.pestis, each characterized by its own emission peak as a marker. Specific and sensitive multiplex analysis for both pathogens has been achieved, down to 10(3) bacteria per ml in the sample.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacillus / classification
  • Bacillus / genetics
  • Bacillus / immunology
  • Bacillus / isolation & purification*
  • Biosensing Techniques
  • Flow Cytometry*
  • Fluorescence Resonance Energy Transfer
  • Immunoglobulin G / chemistry
  • Immunoglobulin G / immunology
  • Light
  • Phylogeny
  • Quantum Dots*
  • Scattering, Radiation
  • Spores, Bacterial / immunology
  • Staining and Labeling
  • Time Factors
  • Yersinia pestis / classification
  • Yersinia pestis / genetics
  • Yersinia pestis / immunology
  • Yersinia pestis / isolation & purification*

Substances

  • Immunoglobulin G