Present address M.D. Feist, Office of Performance Review, Health Resources Services Administration, Seattle, WA, USA. The opinions expressed in this paper are those of the authors and not necessarily those of the US Food and Drug Administration.
Real-time PCR method for Salmonella spp. targeting the stn gene
Article first published online: 31 JUL 2006
Journal of Applied Microbiology
Volume 102, Issue 2, pages 516–530, February 2007
How to Cite
Moore, M.M. and Feist, M.D. (2007), Real-time PCR method for Salmonella spp. targeting the stn gene. Journal of Applied Microbiology, 102: 516–530. doi: 10.1111/j.1365-2672.2006.03079.x
The opinions expressed in this article are those of the authors and not necessarily those of the US Food and Drug Administration.
- Issue published online: 31 JUL 2006
- Article first published online: 31 JUL 2006
- 2005/1086: received 20 September 2005, revised 11 April 2006 and accepted 5 May 2006
- real-time PCR;
- Salmonella bongori;
Aim: To develop a real-time PCR assay for Salmonella spp. targeting the stn gene.
Methods and Results: The presence of stn in the Salmonella bongori genome was found by a BLAST with Salmonella enterica stn sequence. Manual alignment of stn sequences showed that Salm. bongori had 88% sequence identity with Salm. enterica. Two primers (stnL-433 and stnR-561) and a probe (stnP-452) were designed to target conserved regions in stn and meet the requirements of a 5′-nuclease assay. The primers and probe were evaluated against 353 isolates, including 255 Salm. enterica representing 158 serotypes, 14 Salm. bongori representing 12 serotypes and 84 non-Salmonella representing 56 species from 31 genera. All isolates were correctly identified, with the exception of three isolates of Citrobacter amalonaticus, which gave false positives. The limit of detection with cultured Salmonella was 3 CFU per reaction.
Conclusions: The stn real-time PCR method had 100% inclusivity, 96·4% exclusivity and a level of detection of 3 CFU per reaction for cultured Salmonella spp.
Significance and Impact of the Study: The study showed that stn is present in Salm. bongori and is a valid target for both species of Salmonella. The Salmonella s tn real-time PCR is a useful method for identifying Salmonella spp.