Evaluation of the diagnostic value of fluorescent in situ hybridization in a rat model of bacterial pneumonia

Diagn Microbiol Infect Dis. 2013 Aug;76(4):425-31. doi: 10.1016/j.diagmicrobio.2013.04.028. Epub 2013 Jun 7.

Abstract

In severe nosocomial pneumonia, the pathogenic responsibility of bacteria isolated from airways is far from certain, and a lung biopsy is sometimes performed. However, detection and identification of pathogens are frequently unachieved. Here, we developed a protocol for direct visualization of bacteria within the lung tissue using fluorescent in situ hybridization (FISH) in a rat model of Acinetobacter baumannii pneumonia. The reference positive diagnosis of bacterial pneumonia was the presence of pathological signs of pneumonia associated with the proof of bacteria or bacterial PCR products into the parenchyma. By analysis of 122 sets of slices from 26 rats and using the eubacterial probe EUB-338, our results show that FISH reached a sensitivity and a diagnostic accuracy higher than that of optic microscopy (sensitivity: 96% versus 55.4% and diagnostic accuracy: 96.7% versus 66.4%), whereas both approaches had 100% specificity. FISH could be useful especially on negative biopsies from patients with suspected infectious pneumonia.

Keywords: Acinetobacter Baumannii; Diagnostic; Fluorescent in situ hybridization; Lung infection; Rat model; Sensitivity.

MeSH terms

  • Acinetobacter Infections / diagnosis*
  • Acinetobacter Infections / microbiology
  • Acinetobacter baumannii / genetics*
  • Acinetobacter baumannii / isolation & purification
  • Animals
  • Biopsy
  • Fluorescent Dyes
  • In Situ Hybridization, Fluorescence
  • Lung / microbiology*
  • Lung / pathology
  • Oligonucleotides / chemistry
  • Oligonucleotides / genetics
  • Pneumonia, Bacterial / diagnosis*
  • Pneumonia, Bacterial / microbiology
  • Polymerase Chain Reaction
  • Quinolinium Compounds
  • RNA, Bacterial* / analysis
  • RNA, Ribosomal, 16S* / analysis
  • Rats
  • Rats, Sprague-Dawley
  • Sensitivity and Specificity

Substances

  • Alexa fluor 546
  • Fluorescent Dyes
  • Oligonucleotides
  • Quinolinium Compounds
  • RNA, Bacterial
  • RNA, Ribosomal, 16S