In the field of public health, food-borne illnesses are a major concern because it has been estimated that each year 76 million cases occur in the United States causing 5,000 deaths (Suo et al., 2010). In 2008, the Center for Disease Control and Prevention’s FoodNet surveillance program reported over 18 thousand cases of food-borne illnesses occurred in 10 states (Center for Disease Control and Prevention [CDC], 2008). According to estimates from the CDC (2011), the most common food-borne pathogens that maybe seen in the United States are Norvovirus (58%), Clostridium perfringens (10%), Salmonella (11%), Campylobacter spp. (9%), and Staphylococcus aureus (3%). Among the other 9% (not published) include Escherichia coli O157:H7, Shigella spp., Bacillus spp., and other opportunistic pathogens (Prasad & Vidyarthi, 2009). Therefore, accurate and timely detection and identification of food-borne pathogens is crucial for the prevention of food-borne epidemics and the timely treatment of patients with such infections.
Current methods of detection and characterization of food-borne pathogens continues to rely on conventional culturing methods. These culturing methods include enrichment in non-selective and selective media followed by sub-culturing to differential media which can take 2-3 days. Afterwards, characterization of the isolate through biochemical and immunological testing is performed and can take up to 4 days for confirmation (Prasad & Vidyarthi, 2009). It is necessary to quickly identify the pathogen because it may be a matter of life or death for a patient. In all possible food-borne infections, doctors prescribe antibiotics for presumptive treatment of such infections in hope that one of the antibiotics will combat or reduce the numbers of the unknown pathogen until identification is made and the proper treatment initiated (Bodrossy & Sessitsch, 2004).
The emergence of DNA based technologies such as the polymerase chain reaction (PCR) and microarray analysis have been utilized in the rapid detection and identification of many pathogenic bacteria (Mothershed & Whitney, 2006; Versalovic & Lupski, 2002). The expansion of these technologies has significantly enhanced the sensitivity, specificity and the rapid detection of microorganisms (Suo et al., 2010). Microarray technologies have the potential to perform high-throughput detection of multiple pathogens. Recent work with specific oligonucleotide probes suggests that pathogen detection can be performed on a sample that contains a mixed culture of bacteria (Kim et al., 2008). This paper will
Purpose of Research:
Previous work with DNA based technologies to accurately detect and identify human pathogens has been demonstrated. Furthermore, three methods have been utilized: 1) amplification of one or more universal genes (16S rRNA and 23S rRNA) through PCR, 2) amplification of pathogen-specific markers (toxins, virulence factors) using multi-plex PCR and 3) amplification of...