title: Quantitative PCR and Cytology of Bronchoalveolar Lavage Fluid in Dogs with Bordetella bronchiseptica Infection.
abstract: BACKGROUND
  The use of quantitative PCR (qPCR) for detection of Bordetella bronchiseptica in bronchoalveolar lavage fluid (BALF) and demonstration of bacteria adhering to ciliated epithelial cells in BALF or bronchial brushing fluid (BBF) has not been assessed in a series of affected dogs. Coinfections can worsen the clinical severity in bordetellosis, but the specific association with Mycoplasma cynos has not been evaluated.
  
  
  OBJECTIVE
  To assess the utility of culture, qPCR and cytologic examination of cytospin preparations in the diagnosis of bordetellosis in dogs and the influence of coinfection by M. cynos on disease severity.
  
  
  METHODS
  Twenty-four referred dogs with B. bronchiseptica infection and 10 healthy dogs.
  
  
  METHODS
  Retrospective case series. qPCR (B. bronchiseptica and M. cynos) and culture results from BALF were recorded. Cytospin preparations from BALF and BBF were reviewed. qPCR on BALF from 10 healthy dogs were used as negative control.
  
  
  RESULTS
  The BALF culture and qPCR detected B. bronchiseptica in 14/24 and 18/18 dogs, respectively. Coccobacilli were found adhering to ciliated epithelial cells in 20 of the 21 BALF cytologic preparations where epithelial cells were found, and 2/3 BBF cytologic preparations. Quantitative PCR detected a low level of B. bronchiseptica in one healthy dog. The frequency of detection of M. cynos was not significantly different in B. bronchiseptica (9/17 dogs) compared with healthy dogs (2/10 dogs) (P = .09).
  
  
  CONCLUSIONS
  Quantitative PCR detection of B. bronchiseptica in BALF appears to be a useful diagnostic tool. Cytologic examination of BALF or BBF, when positive, allows a rapid and reliable diagnosis.
authors.affiliation: Faculty of Veterinary Medicine, Department of Small Animal Clinical Sciences, Internal Medicine, University of Liège, Liège, Belgium.
authors.forename: A M
authors.initials: AM
authors.lastname: Canonne
authors.affiliation: Faculty of Veterinary Medicine, Department of Small Animal Clinical Sciences, Internal Medicine, University of Liège, Liège, Belgium.
authors.forename: F
authors.initials: F
authors.lastname: Billen
authors.affiliation: Faculty of Veterinary Medicine, Department of Small Animal Clinical Sciences, Internal Medicine, University of Liège, Liège, Belgium.
authors.forename: C
authors.initials: C
authors.lastname: Tual
authors.affiliation: Faculty of Veterinary Medicine, Department of Small Animal Clinical Sciences, Internal Medicine, University of Liège, Liège, Belgium.
authors.forename: E
authors.initials: E
authors.lastname: Ramery
authors.affiliation: Faculty of Veterinary Medicine, Department of Small Animal Clinical Sciences, Internal Medicine, University of Liège, Liège, Belgium.
authors.forename: E
authors.initials: E
authors.lastname: Roels
authors.affiliation: TDDS Ltd., Unit G, The Innovation Centre, University of Exeter, Exeter, UK.
authors.forename: I
authors.initials: I
authors.lastname: Peters
authors.affiliation: Faculty of Veterinary Medicine, Department of Small Animal Clinical Sciences, Internal Medicine, University of Liège, Liège, Belgium.
authors.forename: C
authors.initials: C
authors.lastname: Clercx
mesh_terms: D000818:Animals; D001885:Bordetella Infections / Q000382:microbiology / Q000662:veterinary*; D016950:Bordetella bronchiseptica / Q000472:pathogenicity*; D001992:Bronchoalveolar Lavage Fluid / Q000166:cytology / Q000382:microbiology*; D016022:Case-Control Studies; D004283:Dog Diseases / Q000382:microbiology*; D004285:Dogs; D005260:Female; D008297:Male; D009174:Mycoplasma / Q000472:pathogenicity*; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012141:Respiratory Tract Infections / Q000382:microbiology / Q000662:veterinary*; D012720:Severity of Illness Index
keywords: Bronchial brushing fluid; Clinical pathology; Cytology; Mycoplasma cynos; qPCR