Abstract
Salmonella enterica, Vibrio parahaemolyticus, Escherichia coli and Listeria monocytogenes are human pathogenic bacteria which could be contaminated with many types of food, especially meat products. Testing of food for those pathogens has become routine practice all over the world. However, since the available traditional detection methods are time consuming and labor intensive, there is a need for rapid, sensitive, specific and cost effective detection technique. In this study, a rapid and sensitive single tube Multiplex PCR method for simultaneous detection of those four bacterial pathogens by amplification of target genes gyrB of V. parahaemolyticus, invA of S. enterica, eaeA of E.coli and hly of L. monocytogene was established . Concentration of bacterial cells from food samples for DNA extraction is another problem in testing of food pathogens by PCR techniques. Metal hydroxide immobilization was applied prior to DNA extraction and found to be effective step to overcome this problem. Key words : Food-borne Pathogens; Metal Hydroxide Immobilization; Multiplex PCR DOI: 10.4038/tare.v12i1.1983 Tropical Agricultural Research & Extension 12(1):2009 42-46
Highlights
As consequence of many pre-harvest, harvest and post harvest factors, food and water can be contaminated by human pathogenic microorganisms
The specificity of primers used to detect target gene is very important in the detection of pathogenic organisms from a food sample which could contain a mixture of microorganisms, and this is a major concern in establishment of Multiplex polymerase chain reaction (PCR) protocols
The bacterial DNA extracted from meat samples inoculated with the four bacteria separately were able to produce bands of 285bp, 685bp, 384bp and 482bp for the gene gyrB of Vibrio parahaemolyticus, invA of S. enterica, eaeA of E.coli and hly of L. monocytogene respectively, while the bacterial DNA extracted from meat samples inoculated with a mixture of all of the four bacteria produced the all above four bands (Fig. 1)
Summary
As consequence of many pre-harvest, harvest and post harvest factors, food and water can be contaminated by human pathogenic microorganisms. Salmonella enteritica, Vibrio parahaemolyticus, Escherichia coli and Listeria monocytogenes are four food and water borne pathogenic bacteria causing gastroenteritis of human. Salmonella is one of the major food and water borne pathogenic bacteria which cause the disease called salmonellosis. E. coli can colonize the intestinal tract of cattle and other animals. During slaughtering, they may contaminate the carcass and subsequently be distributed via cut meat or raw meat materials intended for further processing. L. monocytogenes is an opportunistic intracellular pathogen causes the disease called listeriosis. It is considered as an important cause of human food and water borne infections worldwide (Liu 2006)
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