Characterization of rabbit Pasteurella multocida isolates by use of whole-cell, outer-membrane, and polymerase chain reaction typing

Purpose: To characterize Pasteurella multocida isolates from laboratory rab bits using serotyping, sodium dodecyl sulfate-polyacrylamide gel electropho resis 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 w as 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. T en WCP patterns (W1-W10) with one variation (W1a) and 10 GRIP (OM1-OM10) pa tterns were found. Primers M13 phage, modified M13 phage, (GTG),, and (GACA ), generated 7, 9, 5, and 9 fingerprint types, respectively, Combination of WCP, OMP and PCR fingerprint results yielded 39 groups with a discriminati on index of 0.98. The PCR fingerprint results generally indicated clonal as sociation among isolates within geographic locations except for the isolate s from Texas, which varied markedly in PCR fingerprint types. Conclusion: Single primer PCR fingerprinting provided a simple and rapid me ans of typing P. multocida isolates from laboratory rabbits, Combinations o f conventional and molecular typing enhanced differentiation among P, multo cida isolated from rabbits with pasteurellosis.