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. 2017 Aug 1;58(10):4307-4309.
doi: 10.1167/iovs.17-21535.

Peptide Nucleic Acid-Fluorescence In Situ Hybridization for Detection of Staphylococci From Endophthalmitis Isolates: A Proof-of-Concept Study

Nimesh Patel  1 , Darlene Miller  1 , Nidhi Relhan  1 , Harry W Flynn Jr  1
Affiliations

Peptide Nucleic Acid-Fluorescence In Situ Hybridization for Detection of Staphylococci From Endophthalmitis Isolates: A Proof-of-Concept Study

Nimesh Patel et al. Invest Ophthalmol Vis Sci. .

Erratum in

  • Erratum.
    [No authors listed] [No authors listed] Invest Ophthalmol Vis Sci. 2017 Sep 1;58(11):4799. doi: 10.1167/iovs.17-22948a. Invest Ophthalmol Vis Sci. 2017. PMID: 28973336 Free PMC article. No abstract available.

Abstract

Purpose: Rapid identification of pathogens causing endophthalmitis may improve treatment outcomes through early administration of species-specific medication. The current study reports a new molecular application of peptide nucleic acid-fluorescence in situ hybridization (PNA-FISH) with Staphylococcus-specific molecular PNA probes for the potential rapid detection of common pathogens causing endophthalmitis.

Methods: An experimental study was designed to evaluate the proof of concept at the microbiology laboratory of the Bascom Palmer Eye Institute. Stored culture-positive staphylococci endophthalmitis isolates obtained from prior vitreous samples (n = 15), along with broth as negative controls (n = 5) were used. Inoculum was prepared to a final concentration of 1 × 105 colony-forming units/mL to ensure that the isolates were viable. Smears of samples were fixed and hybridized using QuickFISH protocol with probes for Staphylococcus.

Results: With PNA-FISH technique, Staphylococcus aureus was identified in 9 of 10 samples and coagulase-negative staphylococci were identified in 10 of 10 samples. Detection time was 20 minutes.

Conclusions: This study serves a proof of concept using a new microbial detection system with FISH probes, and may have the potential for clinical use in the rapid and accurate identification of isolates from patients with endophthalmitis.

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Figures

Figure
Figure
Vitreous sample as seen under fluorescence microscope (×60 magnification), using PNA-FISH/QuickFISH procedure for rapid detection of pathogens. Staphylococcus aureus is depicted as green-colored cocci in groups and coagulase-negative staphylococci as red-colored cocci in groups.
See this image and copyright information in PMC

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