Journal
JOURNAL OF MICROBIOLOGICAL METHODS
Volume 59, Issue 1, Pages 23-32Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.mimet.2004.05.008
Keywords
bacterial invasion; intracellular bacteria; fluorescence staining; Neisseria gonorrhoeae; Staphylococcus aureus
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Microscopic discrimination between extracellular and invasive, intracellular bacteria is a valuable technique in microbiology and immunology. We describe a novel fluorescence staining protocol, called FITC-biotin-avidin (FBA) staining, which allows the differentiation between extracellular and intracellular bacteria and is independent of specific antibodies directed against the microorganisms. FBA staining of eukaryotic cells infected with Gram-negative bacteria of the genus Neisseria or the Gram-positive pathogen Staphylococcus aureus are employed to validate the novel technique. The quantitative evaluation of intracellular pathogens by the FBA staining protocol yields identical results compared to parallel samples stained with conventional, antibody-dependent methods. FBA staining eliminates the need for cell permeabilization resulting in robust and rapid detection of invasive microbes. Taken together, FBA staining provides a reliable and convenient alternative for the differential detection of intracellular and extracellular bacteria and should be a valuable technical tool for the quantitative analysis of the invasive properties of pathogenic bacteria and other microorganisms. (C) 2004 Elsevier B.V. All rights reserved.
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