Eubacterial PCR - the usefulness of the molecular method in clinical practice

Authors

  • Zala Lužnik University Medical Centre Ljubljana, Slovenia
  • Tjaša Cerar Kišek Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Zaloska str. 4, 1000 Ljubljana, Slovenia
  • Eva Ružić-Sabljić Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Zaloska str. 4, 1000 Ljubljana, Slovenia
  • Manica Müller Premru Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Zaloska str. 4, 1000 Ljubljana, Slovenia
  • Tomaž Jurca Clinic for Infectious Diseases and Febrile Illnesses, University Medical Centre Ljubljana, Japljeva str. 2, 1525 Ljubljana, Slovenia
  • in Janez Tomažič Clinic for Infectious Diseases and Febrile Illnesses, University Medical Centre Ljubljana, Japljeva str. 2, 1525 Ljubljana, Slovenia

DOI:

https://doi.org/10.6016/ZdravVestn.1019

Keywords:

bacterial infections, specimens, cultivation, identification, 16S rRNK PCR

Abstract

Abstract

Background: Isolation and identification of bacterial pathogens enables accurate diagnosis of bacterial infection, allowing rational use of appropriate narrow-spectrum antibiotics. In some cases, the routine bacterial culture can give negative results. In those cases additional use of molecular techniques such as eubacterial (broad-range 16S rRNA) PCR may detect and identify bacterial genetic material.

Methods: Between February 2012 and April 2013 42 specimens from 35 patients, already treated with antimicrobials, were taken and tested by eubacterial PCR in addition to routine microbiological culture. Results: Eubacterial PCR yielded positive result in 21/42 specimens in 18 patients (in three mixed sequences). Therefore, in 15 patients the diagnosis of bacterial infection was obtained with DNA identification and the results were interpreted in accordance to patients’ history, laboratory and image diagnostics. Only 4 specimens were culture-positive.

Conclusions: Although eubacterial PCR enables the identification of any bacterial DNA in clinical specimens, there are some limitations: no information concerning antimicrobial susceptibility of the causative agents, problem of differentiating living from dead bacteria and problem to differentiate contaminants from pathogenic bacteria. The method is also expensive. In the following article recommendations for appropriate and rational use of eubacterial PCR are presented.

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Author Biographies

  • Zala Lužnik, University Medical Centre Ljubljana, Slovenia
    M.D.
  • Tjaša Cerar Kišek, Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Zaloska str. 4, 1000 Ljubljana, Slovenia
    univ. dipl. mikr., Ph.D.
  • Eva Ružić-Sabljić, Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Zaloska str. 4, 1000 Ljubljana, Slovenia
    prof. Eva Ružić-Sabljić, M.D., Ph.D.
  • Manica Müller Premru, Institute of Microbiology and Immunology, Faculty of Medicine, University of Ljubljana, Zaloska str. 4, 1000 Ljubljana, Slovenia
    prof. Manica Müller Premru, M.D., Ph.D.
  • Tomaž Jurca, Clinic for Infectious Diseases and Febrile Illnesses, University Medical Centre Ljubljana, Japljeva str. 2, 1525 Ljubljana, Slovenia
    M.D., MSc
  • in Janez Tomažič, Clinic for Infectious Diseases and Febrile Illnesses, University Medical Centre Ljubljana, Japljeva str. 2, 1525 Ljubljana, Slovenia
    prof. Janez Tomažič, M.D., Ph.D.

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Published

2015-10-19

How to Cite

1.
Eubacterial PCR - the usefulness of the molecular method in clinical practice. ZdravVestn [Internet]. 2015 Oct. 19 [cited 2024 Nov. 2];84(10). Available from: https://vestnik.szd.si/index.php/ZdravVest/article/view/1019