Abstract:
Polymerase Chain Reaction (PCR) based detection methods have received significant
attention in food borne microbial pathogen detection. However, reliability and sensitivity of
these methods are highly depending on the extraction of adequate amount of pure DNA using
appropriate extraction method. Hence, selection of appropriate DNA extraction method is
very important in PCR based detection of microbial pathogens. In this study, the extraction
efficiency of five commonly used DNA extraction methods was evaluated. Salmonella
enterica was used as experimental organism and five extraction methods were tested for their
ability to extract DNA from spiked pork meat samples. Pork meat samples were incubated for
four hours after being added a dilution series (100
– 103 CFU/mL) of Salmonella enterica
culture. Then DNA was extracted from those samples by the five commonly used DNA
extraction methods. Using extracted DNA, fliC gene of Salmonella was amplified by Nested
PCR. Out of those five methods, the modified Fontana and Kapperd methods were found to
be more effective in DNA extraction. In addition, those methods gave high detection
sensitivity of 101CFU/mL in Nested PCR amplification.
