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Direct identification of major Gram-negative pathogens in respiratory specimens by respiFISH® HAP Gram (−) Panel, a beacon-based FISH methodology

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Abstract

Rapid detection of microorganisms in respiratory specimens is of paramount importance to drive the proper antibiotic regimen to prevent complications and transmission of infections. In the present study, the respiFISH® HAP Gram (−) Panel (miacom diagnostics GmbH, Duesseldorf, Germany) for the etiological diagnosis of hospital-acquired pneumonia was compared with the traditional culture method for the detection of major Gram-negative pathogens in respiratory specimens. respiFISH® combined the classical fluorescence in situ hybridization (FISH) technology with fluorescence-labeled DNA molecular beacons as probes. From September 2011 to January 2012, 165 samples were analyzed: the sensitivity and specificity were 94.39 and 87.93 %, respectively. Only six pathogens (3.6 %) were not identified with respiFISH®, while seven specimens (3 %) provided false-positive results. This beacon-based identification shortens the time to result by at least one work day, providing species-level identification within half an hour. Considering the high sensitivity and specificity and the significant time saving, the introduction of bbFISH® assays could effectively complement traditional systems in microbiology laboratories.

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Abbreviations

bbFISH®:

Beacon-based fluorescence in situ hybridization

HAP:

Hospital-acquired pneumonia

VAP:

Ventilation-acquired pneumonia

TAT:

Total turn-around time

CNA:

Columbia nalidixic acid agar

BAL:

Bronchoalveolar lavage

PPV:

Positive predictive value

NPV:

Negative predictive value

PCR:

Polymerase chain reaction

IC:

Interval confidence

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Conflict of interest

All co-authors have no specific conflict of interests.

Author information

Author notes

  1. C. Fontana

    Present address: Clinical Microbiology Laboratories, Foundation Polyclinic Tor Vergata, V.le Oxford 81, 00133, Roma, Italy

Authors and Affiliations

  1. Department of Pathology and Diagnostics, University of Verona, Strada Le Grazie 8, 37134, Verona, Italy

    R. Koncan, V. Favuzzi & M. Ligozzi

  2. O.U. of Microbiology and Virology, Department of Pathology and Diagnostics, Azienda Ospedaliera Universitaria Integrata di Verona, P.le A. Scuro 10, 37134, Verona, Italy

    M. Parisato & G. Lo Cascio

  3. Clinical Microbiology Laboratories, Foundation Polyclinic Tor Vergata, V.le Oxford 81, 00133, Roma, Italy

    C. Sakarikou

  4. Department of Anesthesiological Sciences and Specialistic Surgeries, University of Verona, P.le A. Scuro 10, 37134, Verona, Italy

    G. Stringari

  5. Department of Experimental Medicine and Surgery, “Tor Vergata” University of Rome, via Montpellier 1, 00133, Roma, Italy

    C. Fontana

Authors
  1. R. Koncan
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  2. M. Parisato
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  3. C. Sakarikou
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  4. G. Stringari
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  5. C. Fontana
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  6. V. Favuzzi
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  7. M. Ligozzi
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  8. G. Lo Cascio
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Correspondence to R. Koncan.

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Koncan, R., Parisato, M., Sakarikou, C. et al. Direct identification of major Gram-negative pathogens in respiratory specimens by respiFISH® HAP Gram (−) Panel, a beacon-based FISH methodology. Eur J Clin Microbiol Infect Dis 34, 2097–2102 (2015). https://doi.org/10.1007/s10096-015-2458-y

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  • DOI: https://doi.org/10.1007/s10096-015-2458-y

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