title: Evaluation of the diagnostic value of fluorescent in situ hybridization in a rat model of bacterial pneumonia.
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.
authors.affiliation: Aix Marseille Université, URMITE, UM63, CNRS 7278, IRD 198, Inserm 1095, 13005 Marseille, France.
authors.forename: Thérèse
authors.initials: T
authors.lastname: Atieh
authors.forename: Gilles
authors.initials: G
authors.lastname: Audoly
authors.forename: Sami
authors.initials: S
authors.lastname: Hraiech
authors.forename: Hubert
authors.initials: H
authors.lastname: Lepidi
authors.forename: Antoine
authors.initials: A
authors.lastname: Roch
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Rolain
authors.forename: Didier
authors.initials: D
authors.lastname: Raoult
authors.forename: Laurent
authors.initials: L
authors.lastname: Papazian
authors.forename: Fabienne
authors.initials: F
authors.lastname: Brégeon
mesh_terms: D000151:Acinetobacter Infections / Q000175:diagnosis* / Q000382:microbiology; D040981:Acinetobacter baumannii / Q000235:genetics* / Q000302:isolation & purification; D000818:Animals; D001706:Biopsy; D005456:Fluorescent Dyes; D017404:In Situ Hybridization, Fluorescence; D008168:Lung / Q000382:microbiology* / Q000473:pathology; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics; D018410:Pneumonia, Bacterial / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction; D011806:Quinolinium Compounds; D012329:RNA, Bacterial / Q000032:analysis; D012336:RNA, Ribosomal, 16S / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D012680:Sensitivity and Specificity
chemical_list: C481052:Alexa fluor 546; D005456:Fluorescent Dyes; D009841:Oligonucleotides; D011806:Quinolinium Compounds; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S
keywords: Acinetobacter Baumannii; Diagnostic; Fluorescent in situ hybridization; Lung infection; Rat model; Sensitivity