Accelerated identification of Staphylococcus aureus from blood cultures by a modified fluorescence in situ hybridization procedure.

Accelerated identification of Staphylococcus aureus from blood cultures by a modified fluorescence in situ hybridization procedure.

Journal of medical microbiology (2009-10-03)

Sven Poppert, Melanie Riecker, Nele Wellinghausen, Hagen Frickmann, Andreas Essig

PMID19797464

ABSTRACT

This study evaluated fluorescence in situ hybridization (FISH) for rapid identification of Staphylococcus aureus and coagulase-negative staphylococci (CoNS) directly from blood cultures. Initially, 360 blood cultures containing Gram-positive cocci were investigated by a previously described microwave-FISH procedure: 44/49 (89.8 %) S. aureus and 298/299 (99.7 %) CoNS were correctly identified. Because FISH proved useful and reliable but handling was found to be inconvenient, the method was modified by employing a recently developed slide chamber. This reduced the time required from 60 to 30 min. The simplified execution allowed integration of the method into the workflow of a routine laboratory without difficulty. The modified method proved to be highly reliable, identifying 37/37 (100 %) S. aureus and 169/172 (98.2 %) CoNS directly from blood cultures.

MATERIALS

Product Number

Brand

Product Description

MBD0047

Sigma-Aldrich

Enterococcus faecium FISH probe – ATTO 488, Probe for fluorescence in situ hybridization (FISH), 20µM in water

MBD0029

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Porphyromonas gingivalis FISH probe - Cy3, Probe for fluorescence in situ hybridization (FISH),20 μM in water

MBD0032

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Eubacteria FISH probe - ATTO488, Probe for fluorescence in situ hybridization (FISH), 20 µM in water

MBD0035

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Bacterial negative control FISH probe - Cy3, Probe for fluorescence in situ hybridization (FISH),20 μM in water

MBD0040

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Proteus FISH probe - Cy3, Probe for fluorescence in situ hybridization (FISH),20 μM in water

MBD0049

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Escherichia coli FISH probe -ATTO488, Probe for fluorescence in situ hybridization (FISH), 20µM in water

MBD0048

Sigma-Aldrich

Enterococcus faecium FISH probe - Cy3, Probe for fluorescence in situ hybridization (FISH), 20µM in water

MBD0050

Sigma-Aldrich

Akkermansia muciniphila FISH probe-ATTO488, Probe for fluorescence in situ hybridization (FISH), 20µM in water

MBD0041

Sigma-Aldrich

Akkermansia muciniphila FISH probe - Cy3, Probe for fluorescence in situ hybridization (FISH),20 μM in water

MBD0059

Sigma-Aldrich

Alphaproteobacteria FISH probe-Cy3, Probe for fluorescence in situ hybridization (FISH), 20 μM in water

MBD0033

Sigma-Aldrich

Eubacteria FISH probe - Cy3, Probe for fluorescence in situ hybridization (FISH)

MBD0028

Sigma-Aldrich

Porphyromonas gingivalis FISH probe - ATTO488, Probe for fluorescence in situ hybridization (FISH)

MBD0034

Sigma-Aldrich

Bacterial negative control FISH probe - ATTO488, Probe for fluorescence in situ hybridization (FISH),20 μM in water

MBD0042

Sigma-Aldrich

Escherichia coli FISH probe - Cy3, Probe for fluorescence in situ hybridization (FISH)