Broad-range (pan) Salmonella and Salmonella serotype typhi-specific real-time PCR assays: potential tools for the clinical microbiologist.
We describe broad-range salmonellae (ie, Salmonella) and Salmonella serotype Typhi-specific LightCycler (Roche Diagnostics, Indianapolis, IN) real-time polymerase chain reaction assays. We validated these with a battery of 280 bacteria, 108 of which were salmonellae representing 20 serotypes. In addition, 298 isolates from 170 clinical specimens that were suspected to possibly represent Salmonella were tested with the pan- Salmonella assay. Finally, the pan-Salmonella assay also was used to test DNA extracts from 101 archived, frozen stool specimens, 55 of which were culture-positive for salmonellae. Both assays were 100% sensitive and specific when cultured isolates of the battery were tested. The pan- Salmonella assay also characterized correctly all salmonellae on the primary isolation agar and was 96% sensitive (53/55) and 96% specific (49/51) when nucleic acid extracts from direct stool specimens were tested. These assays represent potential tools the clinical microbiologist could use to screen suspect isolates or stool specimens for Salmonella.
Farrell
John J
JJ
Department of Infectious Diseases, University of, Illinois/OSF Infectious Diseases, Peoria, IL, USA.
Doyle
Laura J
LJ
Addison
Rachel M
RM
Reller
L Barth
LB
Hall
Geraldine S
GS
Procop
Gary W
GW
D004269:DNA, Bacterial / Q000032:analysis*; D005243:Feces / Q000382:microbiology; D006801:Humans; D008828:Microbiological Techniques / Q000379:methods*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012475:Salmonella / Q000145:classification / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D012703:Serotyping
D004269:DNA, Bacterial