Identifying intracellular Staphylococcus aureus in chronic rhinosinusitis: A direct comparison of techniques

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The emerging concept of intracellular pathogens such as Staphylococcus aureus playing a role in chronic rhinosinusitis (CRS) has led to the development of numerous imaging techniques for their identification. Traditional methods of bacterial culture are not effective at localizing bacteria to the surface or within tissue samples. The aim of this study was to develop and validate a novel imaging technique using confocal scanning laser microscopy (CSLM) coupled with a fluorescence in situ hybridization (FISH) probe and nucleic acid counterstain (propidium iodide [PI]) that allows for simultaneous analysis of S. aureus intracellular status and surface biofilm within whole mucosal samples.


A prospective study was performed including 17 patients undergoing endoscopic sinus surgery for CRS. Tissue samples were analyzed with both CSLM-FISH/PI and immunohistochemistry (IHC) for intracellular S. aureus status.


Using CSLM-FISH/PI intracellular S. aureus was identified in 9/17 (47%) patients and in 7/17 (39%) using IHC. Surface biofilm can be identified with CSLM-FISH/PI in the same piece of tissue; however, deeper imaging to the submucosa is impossible. IHC showed submucosal bacteria in three patients.


Both CSLM-FISH/PI and IHC are complementary techniques that can be used to identify intracellular S. aureus. CSLM-FISH/PI allows for the simultaneous detection of intracellular status and surface biofilm within the tissue analyzed. IHC has a role in the identification of intracellular and submucosal S. aureus within these tissues. Additional investigation is required to identify the true pathogenic nature of intracellular organisms as well as any relationship to surface biofilm status.

Keywords: Chronic rhinosinusitis; Staphylococcus aureus; fluorescence in situ hybridization; imaging; immunohistochemistry; intracellular

Document Type: Research Article


Affiliations: Department of Surgery–Otolaryngology Head and Neck Surgery, University of Adelaide, Adelaide, South Australia, Australia

Publication date: November 1, 2012

More about this publication?
  • The American Journal of Rhinology & Allergy, is a peer reviewed, scientific publication committed to expanding knowledge and publishing the best clinical and basic research within the fields of Rhinology & Allergy. Its focus is to publish information which contributes to improved quality of care for patients with nasal and sinus disorders. Its primary readership consists of otolaryngologists, allergists, and plastic surgeons. Published material includes peer-reviewed original research, clinical trials and review articles.

    Articles marked "F" offer free full text for personal noncommercial use only.

    Previously published as American Journal of Rhinology, the journal is indexed in Thomson Reuters Web of Science and Science Citation Index, plus the National Library of Medicine's PubMed service.
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