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. 2005 Sep;43(9):4855-7.
doi: 10.1128/JCM.43.9.4855-4857.2005.

Rapid identification of Staphylococcus aureus in blood cultures by a combination of fluorescence in situ hybridization using peptide nucleic acid probes and flow cytometry

Hanna Hartmann  1 , Henrik Stender, Andrea Schäfer, Ingo B Autenrieth, Volkhard A J Kempf
Affiliations

Rapid identification of Staphylococcus aureus in blood cultures by a combination of fluorescence in situ hybridization using peptide nucleic acid probes and flow cytometry

Hanna Hartmann et al. J Clin Microbiol. 2005 Sep.

Abstract

Fluorescence in situ hybridization (FISH) using peptide nucleic acid probes (PNAs) allows the identification of Staphylococcus aureus from human blood culture samples. We present data revealing that the combination of PNA FISH and flow cytometry is a possible approach for the noncultural identification of staphylococci in blood cultures.

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Figures

FIG. 1.
FIG. 1.
Identification of S. aureus in blood culture samples by PNA FISH using flow cytometry. (A and B) Blood cultures were spiked with S. epidermidis (ATCC 12228) or S. aureus (ATCC 25923). PNA FISH was performed in liquid hybridization buffer and analyzed microscopically (panel A) revealing hybridization of the S. aureus-specific PNA probe (counterstaining with DAPI [4′,6′-diamidino-2-phenylindole]; scale bar, 4 μm). Panel B consists of flow cytometric histograms showing the intensities of green fluorescence (FL1-H) on the x axes and the cell counts on the y axes. (C and D) Identification of staphylococci in blood culture samples spiked with clinical isolates of staphylococci. After growth detection by using BACTEC 9240, liquid FISH and flow cytometric analysis were performed. Data analysis revealed identifications of S. aureus (panel C) due to the shift in green fluorescence (arrow, to the right of the dotted line) and of CoNS (panel D) due to the missing green fluorescence (to the right of the dotted line). Means are mean values of fluorescence intensities.
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