TITLE

GREITAS SALMONELLA TYPHIMURIUM IR SALMONELLA CHOLERAESUIS NUSTATYMAS KIAULIŲ FEKALIJŲ MĖGINIUOSE MODIFIKUOTA LIZDINE POLIMERAZĖS GRANDININE REAKCIJA

AUTHOR(S)
Stankevičus, Arūnas; Čepulis, Rytis; Sajūtė, Kristina; Stankevičienė, Marija
PUB. DATE
December 2005
SOURCE
Veterinarija ir Zootechnika;2005, Vol. 32 Issue 54, p32
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
Public health implications of Salmonella infections in animals and products of thereof enforce strengthening of the surveillance and search for efficient diagnostic tools. Conventional Salmonella detection takes several days. Polymerase chain reaction (PCR) or conventional nested PCR tests have been developed to shorten analysis time but still can generate contaminants due to complex manipulations and can be hampered by inhibitors of PCR present in the tested sample. To overcome those problems and shorten analysis time we have developed one-tube nested PCR assay based on the detection of Salmonella-specific invA gene using well defined primer sets. Swine feaces spiked with S. typhimurium and S. choleraesuis were used as a sample model. The assay was able to detect 150 CFU/g feaces compared to 15 CFU/ml in saline. Simultaneously, the detection level of the applied single PCR tests as well as conventional nPCR reached 150 and 15 CFU/ml of saline and, respectively, 15 × 107 and 15 × 10² CFU/g of swine feaces. The sensitivity of the test was increased up to 15 CFU/g using enrichment in buffered peptone water for 4 hours. No serovar-dependent differences were observed. It was concluded that 4-step procedure including 4-hour nonselective pre-enrichment, DNA extraction, PCR amplification and electrophoretic detection of known molecular weight product (283 bp) can be used as a screening test for Salmonella detection in swine feaces.
ACCESSION #
19414051

 

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