Evaluation of BIOTECH PCR-based detection kit for Salmonella in foods, water and feeds
Ramirez, T.J. | Perez, M.T.M. | Mercado, M.A. | Sedano, S.A. | Sapin, A.B. | Calapardo, M.R. | Garcia, M.R.C. | Tambalo, R.D. | Guerra, M.A.R.V. | Mercado, S.M. (Philippines Univ. Los Banos, 4031 College, Laguna (Philippines). National Inst. of Molecular Biology and Biotechnology)
The current study compares a rapid PCR-based detection method developed at BIOTECH with the traditional cultural approach involving enrichment step under plating and selective media. The BIOTECH PCR-based Salmonella detection kit consisted of reaction tubes containing a selected primer for Salmonella, PCR buffer, deoxynucleotides, and magnesium chloride and taq polymerase enzyme. The detection protocol includes short enrichment period to allow multiplication of Salmonella, DNA extraction and amplification of DNA in a thermal cycles followed by a simple gel electrophoresis procedure. The presence of 0.9 Kb product indicates the presence of Salmonella in the samples. Various samples of foods, water and feeds were monitored for the presence of Salmonella following the AOAC procedure and the BIOTECH protocol involving the PCR-based kit. Validation of the detection kit was done both on naturally contaminated and artificially spiked samples. Known number of Salmonella cells were spiked on both the conventional and the BIOTECH PCR-based kit can detect Salmonella in the artificially-inoculated food samples. Lower agreement values were obtained on naturally contaminated samples due to the low number of Salmonella cells present after enrichment period. The minimum Salmonella cells load that is detectable by the BIOTECH kit is about 10E4 cells per ml. A more intense band was observed in samples when the cell load is higher than this level
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Este registro bibliográfico ha sido proporcionado por University of the Philippines at Los Baños