Submitted to: CRC Press
Publication Type: Book / Chapter
Publication Acceptance Date: 3/31/2007
Publication Date: 9/1/2007
Citation: Fratamico, P.M., Kawasaki, S. 2007. Applications of the polymerase chain reaction (pcr) for detection, identification, and typing of food-borne microorganisms. CRC Press, New York. Microbial Food Contamination, C.Wilson 9ed.) 2nd edition. p.213-254
Technical Abstract: Public awareness of microorganisms transmitted by food, which pose a severe threat to human health, has increased dramatically in recent years. Some of the agents, which have been responsible for numerous cases and outbreaks of food-borne illness and also many deaths, are Salmonella spp., Listeria monocytogenes, Escherichia coli O157:H7 and other Shiga toxin-producing E. coli, Yersinia enterocolitica, Clostridium perfringens, Giardia, Cryptosporidium, Cyclospora, hepatitis A virus, and noroviruses. Thus, there is a need for rapid, sensitive, and reliable methods for detection of food-borne pathogens and for investigating cases and outbreaks of illness caused by these agents. Advancements in technology have resulted in the development of various types of rapid methods for detection, identification, and enumeration of food-borne pathogens. In addition to providing results in a shorter length of time than conventional culture-based methods, rapid assays are often more sensitive, specific, and more accurate than classical methods. Rapid methods include miniaturized biochemical kits and antibody- and nucleic acid-based assays. The PCR is a nucleic acid amplification technique, which has become a very useful tool for rapid detection, identification, and typing of pathogenic microorganisms both in the clinical setting and in food and environmental microbiology laboratories. The PCR is a rapid technique, with both high sensitivity and specificity. In recent years, the PCR has advanced from end-point detection to detection of product while the reaction is occurring. This is referred to as “real-time” PCR, and the technique can be used to quantify target DNA. Furthermore, there is currently much interest in the development of rapid diagnostic systems that have the capability to link sample processing, preparation, and simultaneous detection of multiple pathogens and new threats, including potential bio-threat agents. Use of rapid methods, particularly techniques based on the PCR, are important to enhance the safety of the food supply.