Serotyping and ribotyping of Salmonella using restriction enzyme PvuII.
The subtyping and identification of bacterial pathogens throughout food processing and production chains is useful to the new hazard analysis critical control point-based food safety plans. Traditional manual serotyping remains the primary means of subtyping Salmonella isolates. Molecular biology techniques, however, offer the promise of more rapid and sensitive subtyping of Salmonella. This study evaluates the potential of restriction enzyme PvuII, followed by probing with the rRNA operon from Escherichia coli, to generate serotype-specific DNA fingerprints. A total of 32 identified serotypes were found with an overall agreement in 208 of the 259 (80%) isolates tested between U.S. Department of Agriculture serotype identification and riboprint serotype identification. Many of the isolates that did not correlate were serotype identified as Salmonella Montevideo, which indicates that for this serotype, there are multiple ribotypes. When Salmonella Montevideo isolates were not included, the ribotype identification agreed with serotyping in 207 of the 231 (90%) isolates. The primary outcome of any ribotyping procedure is to give distinct ribotype patterns. This extensive poultry epidemiological study demonstrates that, in addition to ribotype patterns, the identification of isolates to known serotypes provides the investigator with additional information that can be more useful than traditional epidemiology and isolate identification studies.
Bailey
J S
JS
U.S. Department of Agriculture, Agricultural Research Service, Athens, Georgia 30604-5677, USA. jsbailey@saa.ars.usda.gov
Fedorka-Cray
P J
PJ
Stern
N J
NJ
Craven
S E
SE
Cox
N A
NA
Cosby
D E
DE
D000818:Animals; D015252:Deoxyribonucleases, Type II Site-Specific; D005516:Food Microbiology; D011200:Poultry / Q000382:microbiology*; D012335:RNA, Ribosomal / Q000032:analysis*; D015203:Reproducibility of Results; D021521:Ribotyping / Q000379:methods*; D012475:Salmonella / Q000145:classification*; D012680:Sensitivity and Specificity; D012703:Serotyping / Q000379:methods*; D013997:Time Factors
D012335:RNA, Ribosomal; C025630:CAGCTG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific