Characterization of rabbit Pasteurella multocida isolates by use of whole-cell, outer-membrane, and polymerase chain reaction typing.
OBJECTIVE
To characterize Pasteurella multocida isolates from laboratory rabbits using serotyping, sodium dodecyl sulfate-polyacrylamide gel electrophoresis of whole-cell proteins (WCPs) and outer-membrane proteins (OMPs), and polymerase chain reaction (PCR) fingerprinting.


METHODS
Fifty isolates were obtained from five sources: ATCC (1), Oklahoma (4), Michigan (9), Minnesota (7), and Texas (29). The PCR fingerprinting was conducted using two minisatellite probes for M13 and a modified M13 core sequence and two microsatellite probes--(GTG)5 and (GACA)4.


RESULTS
Forty-five isolates were serogroup A, and five were serogroup D. Ten WCP patterns (W1-W10) with one variation (W1a) and 10 OMP (OM1-OM10) patterns were found. Primers M13 phage, modified M13 phage, (GTG)5, and (GACA)4 generated 7, 9, 5, and 9 fingerprint types, respectively. Combination of WCP, OMP, and PCR fingerprint results yielded 39 groups with a discrimination index of 0.98. The PCR fingerprint results generally indicated clonal association among isolates within geographic locations except for the isolates from Texas, which varied markedly in PCR fingerprint types.


CONCLUSIONS
Single primer PCR fingerprinting provided a simple and rapid means of typing P. multocida isolates from laboratory rabbits. Combinations of conventional and molecular typing enhanced differentiation among P. multocida isolated from rabbits with pasteurellosis.
Dabo
S M
SM
Department of Anatomy, Pathology, and Pharmacology, College of Veterinary Medicine, Oklahoma State University, Stillwater 74078-2007, USA.
Confer
A W
AW
Montelongo
M
M
Lu
Y S
YS
D000818:Animals; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis*; D001426:Bacterial Proteins / Q000032:analysis*; D016172:DNA Fingerprinting; D004269:DNA, Bacterial / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D018895:Microsatellite Repeats; D018598:Minisatellite Repeats; D016979:Pasteurella multocida / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D011817:Rabbits / Q000382:microbiology*; D012703:Serotyping
D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D004269:DNA, Bacterial