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Detection of Salmonella in Alfalfa Sprouts by Reverse
Transcriptase Polymerase Chain Reaction
2002
FDA Science Forum FDA:
Building a Multidisciplinary Foundation February
20-21, 2002 Washington
Convention Center, Washington, DC Poster
Abstracts, Board T-06 Grisselle Martinez, Karlygash M. Yermukan, ORA, FDA,
Alameda, CA A fast and reliable method for the detection of live Salmonella
cells in alfalfa sprouts using reverse transcriptase PCR was developed during
this study. Total RNA was isolated from sprout samples spiked with Salmonella.
The ST11 and ST15 primers, Aabo et al. were used to amplify a 429 bp-long
fragment during RT-PCR. The resulting DNA fragment was isolated and sequenced.
This method was also tested using sprouts homogenate spiked with Salmonella and
naturally contaminated sprouted seeds cells. The detection limit was ~103
- 104 cells. The estimated time of completion for the rinse, RNA
isolation, RT-PCR and gel electrophoresis was 6 hours for 5-10 samples. The
advantages of this method over the currently used rapid method, are: a) the
detection of live (vs. non-viable) cells, b) the procedure is completed in a
significantly shorter period of time. |