Single primer polymerase chain reaction fingerprinting for Pasteurella multocida isolates from laboratory rabbits

Objective-To evaluate a rapid polymerase chain reaction (PCR) fingerprintin g technique for discriminating among Pasteurella multocida isolates from la boratory rabbits. Sample Population-33 P multocida isolates from rabbits with clinical pasteu rellosis. Procedure-PCR assays were conducted with 2 minisatellites (core sequence an d modified core sequence of phage M13) and 2 microsatellites ([GTG](5) and [GACA](4)). Each bacterium was assigned to a PCR type for each of the prime rs used. Boiled bacterial extracts and purified genomic DNA were compared b y use of PCR assays for phage M13 and (GACA)4. Plasmids were isolated from each bacterium, and their influence on PCR fingerprint was determined, usin g boiled extracts as a DNA source, Results-M13 core sequence and M13 modified core sequence yielded 5 and 8 PC R types, respectively. The microsatellites (GTG)(5) and (GACA)(4) yielded 4 and 9 PCR fingerprint types, respectively. Fingerprint patterns obtained b y use of isolated DNA differed from those obtained by use of boiled extract s, although discrimination among P multocida isolates was similar. The pres ence or absence of plasmids did not affect PCR fingerprints. Conclusion-Single primer PCR fingerprinting with minisatellite and microsat ellite primers is an efficient and reproducible method for the discriminati on of P multocida isolates from rabbits and can be performed directly, usin g boiled bacterial extracts as a source of template, although more bands we re obtained from pure genomic DNA.