Simultaneous Detection of Nine Foodborne Pathogenic Bacteria Using High-resolution Melting Analysis
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Abstract:
Foodborne diseases have become a global food safety concern. Common bacterial pathogens involved in food poisoning include pathogenic Escherichia coli (especially enterohemorrhagic Escherichia coli O157:H7), Salmonella, Shigella, pathogenic Vibrio (including Vibrio cholerae and Vibrio parahaemolyticus), Staphylococcus aureus, Listeria monocytogenes, Campylobacter jejuni, and Cronobacter sakazakii. Species-specific detection for one type of pathogen cannot meet the requirements of modern society for multiple-pathogen detection. In order to establish a method that can simultaneously detect the above-mentioned nine pathogenic bacteria in food, a multiple high-resolution melting (HRM) real-time polymerase chain reaction (PCR) assay for the simultaneous detection of multiple foodborne pathogens was established, using the specific genes of the nine aforementioned bacteria as target genes and a novel pair of universal primers. The results showed that this multiple HRM-real time PCR detection system could effectively detect and distinguish these nine common pathogenic bacteria in food via a two-tube PCR reaction, with good specificity and sensitivity.
