Verification studies on the use of BIOTECH PCR-based detection kit for Salmonella in animal feeds
2002
Ramirez, T.J.,University of the Philippines Los Baños, College, Laguna (Philippines). National Institute of Molecular Biology and Biotechnology | Sedano, S.A.,University of the Philippines Los Baños, College, Laguna (Philippines). National Institute of Molecular Biology and Biotechnology | Guerra, M.A.R.V.,University of the Philippines Los Baños, College, Laguna (Philippines). National Institute of Molecular Biology and Biotechnology | Garcia, M.R.C.,University of the Philippines Los Baños, College, Laguna (Philippines). National Institute of Molecular Biology and Biotechnology | Mercado, S.M.,University of the Philippines Los Baños, College, Laguna (Philippines). National Institute of Molecular Biology and Biotechnology | Perez, M.T.M.,University of the Philippines Los Baños, College, Laguna (Philippines). National Institute of Molecular Biology and Biotechnology
A detection kit employing polymerase chain reaction (PCR) technology was developed for Salmonella. Its application as Salmonella detection kit was investigated using different kinds of feeds, both in naturally contaminated and artificially spiked feeds samples. The kit was compared and validated against standard/traditional cultural assay involving enrichment step and plating in selective media. The BIOTECH Salmonella detection protocol included short enrichment period that allowed multiplication of Salmonella, DNA extraction and amplification of DNA in a thermal cycler followed by a simple gel electrophoresis procedure. Both then conventional and the BIOTECH PCR-based kit can detect the microorganism in Salmonella-inoculated feed samples, with 100 percent agreement value. Slightly lower agreement value (96%) was obtained on natural-contaminated samples due to low number of Salmonella cells present even after enrichment period. Standard cultural method required 5-7 days to confirm the presence and identity of salmonella, while the PCR-based kit reduced the detection time to less than 28 hours.
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