Detection of Cilia-Associated Respiratory Bacillus in Experimentally and Naturally Infected Mice and Rats by the Polymerase Chain Reaction.

Abstract

Cilia-associated respiratory (CAR) bacillus was detected by means of the reverse transcription (RT)-polymerase chain reaction (PCR), and the results were compared with those of indirect immunofluorescence test (IFAT) for the detection of the organism. In the experimental infections, 15 mice were in contact with mice previously inoculated with CAR bacillus. Three mice each were tested at days 3, 5, 7, 12 and 20 postexposure. On day 3 postexposure, CAR bacillus was detected in oral swab samples from all 3 mice by RT-PCR, but was not detected in any sampling sites from the mice by IFAT. Total numbers of positive samples from nasal, oral and tracheal swabs obtained through the test were 6/15, 14/15 and 8/15, respectively, by RT-PCR, and 2/15, 6/15 and 3/15, respectively by IFAT. For the detection of CAR bacillus in samples from 52 rats, 34 serum antibody negative rats by enzyme-linked immunosorbent assay were also negative by RT-PCR and IFAT except for one sample from the oral cavity, and all serum antibody positive rats were positive for the organism by RT-PCR but it could not be detected in five of them by IFAT. By means of RT-PCR, no differences in the positive rates depending on sampling sites were observed except in one rat. The RT-PCR was found to be a specific, highly sensitive and reliable procedure for detecting CAR bacillus in mice and rats. The oral cavity was the most suitable site for the diagnosis of the early stage of this infection by RT-PCR.

Identification Procedure for Pasteurella Pneumotropica in Microbiologic Monitoring of Laboratory Animals

“…Each PCR product was sequenced by an automated sequencer (310 Genetic Analyzer; Applied Biosystems Japan, Tokyo). The data were analyzed using a software package, DNASIS (Hitachi Software Engineering, Tokyo), and compared with the sequences of P. pneumotropica biotype Jawetz (M75083) and Heyl (AF012090), P. dagmatis (M75051), P. aerogenes (M75048), P. multocida subsp multocida (M35018) and Actinobacillus muris (AF024526) in the Gen Bank database for homology analysis [4,6,7,17].…”

Biology and Diseases of Mice

“…An ELISA for serological detection of infection is available (Shoji et al, 1988), and PCR-based diagnostics also have been developed (Goto et al, 1995). Histologic assessment of infection requires the use of Warthin-Starry or similar stains to visualize argyrophilic bacilli adherent to the apical membranes of bronchial respiratory epithelium (Fig.…”

Cilia-associated respiratory bacillus infection in rats in New Zealand

“…Electron microscopy, immunohistochemistry (Oros et al 1996), polymerase chain reaction (Goto et al 1995) and enzyme-linked immunosorbent assay (Shiji et al 1988) techniques have also been used. The bacillus is fastidious and was previously only cultivated in embryonated chicken eggs and cell culture, though artifi cial media are now available but not widely used (Schoeb et al 1993).…”