Electronic Resource
Oxford, UK
:
Blackwell Publishing Ltd
Journal of rapid methods and automation in microbiology
10 (2002), S. 0
ISSN:
1745-4581
Source:
Blackwell Publishing Journal Backfiles 1879-2005
Topics:
Biology
Notes:
A DNA binding fluorescence method based on polymerase chain reaction (PCR) products was evaluated for rapid detection of Salmonella Typhimurium in poultry products. Wash water samples of chicken carcasses and ground turkey were inoculated with S. Typhimurium to obtain final concentrations of 10° - 105 CFU/mL. One mL of each sample was used to get the DNA template and 5 μL of the sample template was added into 25 μL of SYBR Green PCR Master Mix and two specific Salmonella ompC gene primers. The negative control was the same except 5 μL of each wash solution was added instead of 5 μL sample template. The reaction was carried out in a thermocycler. Finally, the fluorescence signal of each PCR product was measured using a fluorometer. The PCR products were also confirmed by ethidium bromide agarose gel, and the DNA concentrations of the PCR products were measured by a filter fluorescence photometer. The results showed that when bacterial cells increased from 0 to 2 CFU/mL, the fluorescence signal increased significantly. The PCR-based fluorescence method could detect the target bacteria in minutes after PCR amplification compared to hours by gel electrophoresis and also could be done at an earlier time during PCR amplification. The detection limit of this method for S. Typhimurium in the poultry samples was 2 CFU/mL without any enrichment.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1111/j.1745-4581.2002.tb00014.x
Permalink
|
Location |
Call Number |
Expected |
Availability |
