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KMID : 0665220040170030254
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Korean Journal of Food and Nutrition
2004 Volume.17 No. 3 p.254 ~ p.259
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Use of RAPD-PCR(Random Amplified Polymorphic DNA-Polymerase Chain Reaction) Method for a Detection of Pathogenic Listeria monocytogenes
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Shin Eon-Hwan
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Abstract
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Rapid detection of foodbome pathogens is becoming increasingly important. The requirement for faster, more reliable tests has lead to the development of a wide range of rapid methods. Among these methods, the use of systems based on nucleic acid based detection has been increasing since they offer advantages of reduction in test time and more reliable detection or identification. Random Amplification Polymorphic DNA(RAPD) method has been used to fingerprint foodbome microorganisms; Listeria monocytogenes. In this study, 10-mer primer OPG-13(5′-CTCTCCGCCA-3′) was used to generate RAPD-PCR for detection of pathogenic L. monocytogenes of Listeria spp. Among 20 primers tested, OPG-13 showed on acceptable result for the differentiation of a pathogenic Listeria from non-pathogenic microorganisms. Pathogenic Listeria, L. monocytogenes(ATCC 15313, 19111, 19112, 19113) showed two bands for 700 bp and 1,500 bp while non-pathogenic bacteria, L. ivanovii, L. grayi, L. murrayi, L. innocua, L. welshimeri, and L. seeligeri had only one band sizing from 2,000 to 2,300 bp. This RAPD method proved to be a valuable to gain important information on sources of pathogenic bacteria in food industry.
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KEYWORD
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PCR, RAPD, Listeria monocytogenes
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