Abstract:
Introduction: Foodborne pathogens are a major cause of disease and death among the global population. illnesses related to contaminated food may vary from person to person from temporary to long-term complications. Nevertheless, therapid rise of multidrug-resistant bacteria worldwide with a declination in antibiotics developments and production make bacteriophages an attractive tool to overcome bacterial resistance. Bacteriophages have become widely recognized for several potential applications in food industry. Theyrepresent an ideal tool for a rapid and sufficient diagnostic assays with great potentials in controlling the spread of harmful pathogens. Their abundance in nature of and high specificity against a specific host bacterium allow them to eradicate, prevent foodborne illness and recalls and provide safe food to consumers.
Purpose: The purpose of this topic is to shed a light on the existing phage based application such as immobilization of phages and using them as a biosensor for foodborne pathogen detection in food.
Methods: In our research, a phage capture-amplification assay based on the phage immobilization on bioactive paper were used. Experiments started with as isolation of very specific phage against one or more of Non O157:H7 E. coliin food. Isolated phages are screened and made to undergo a variety of phenotypic and genotypic characterisationto make sure that they meet the desirable requirement For example host range experiment , efficiency of plating, phage adsorption, growth curve of phage, whole genome sequencing, immobilization phage into colorlok paper (dipstick approach) using electrostatic properties of phages and the surface. The data were collected from three independents trials where the averages and standard deviations were determined. Detection limits were calculated and compared using an Independent-Samples T-Test using IBM SPSS.
Results: The result have statistically significant differences (p<0.05) in the detection of E. coli O45:H2. The cycle threshold (Ct) values were averaged for each concentration and compared to the average Ct values for the incubated control paper without phage. Using phages as biosensor enabled the detection of as few as 10 CFU/mL of the Big Six Shiga Toxin producing E. coli strains in both TSB media and ground beef using both a plaque assay and real-time PCR to detect phage progeny.
Significance: These data suggest that the immobilization of bacteriophages onto paper offers a promising approach for the detection of foodborne pathogens. The bacteriophage dipstick assay had the ability to detect as few as 10 cells of E. coliO45:H2 in only 8 hours in both TSB and ground beef.
