CD Genomics Perspective: DNA-Based Technologies for Detection of Foodborne Microorganisms
Food-borne diseases are complex in nature and appear to pose serious public health issues worldwide. In humans, certain pathogens cause illnesses that are spread by different food products. Consumers, businesses, and regulatory authorities have been very concerned about the microbiological quality of food. Contamination can occur by air, water, soil contamination and at different points during the growing, harvesting, processing, and preparation of food crops. The prevalence of bacteria in food causes problems with food poisoning. Microbiological research is needed to determine the consistency and protection of food. Microbe development can lead to organoleptic changes in food that make it intolerable to the user. An imperative concern in the scientific community is the fast detection and diagnosis of microorganisms. There are two major purposes of food microbiological testing: to assess the disappearance of pathogens or their contaminants in order to ensure food protection, and to list total or microbial load indicators in order to track the efficacy of hygienic handling and to check the consistency of the substance and the durability of the shelf life. We list some commonly used foodborne microorganism research methods based on DNA technology as below.
Restriction Fragment Length Polymorphism
Restriction Fragment Length Polymorphism (RFLP) is a very basic technique using genomic DNA-specific restriction enzyme digestion. It is used to compare the number and size (mass) of fragments formed by cutting restriction endonucleases at a given target DNA recognition site. Electrophoretic isolation investigates the resultant DNA fragments. Proof of evolving DNA sequences is the presence, lack, or changes in the mass of the resultant DNA fragments. For the discrimination of bacteria at the species level, this technique requires pure culture. RFLP was used in conjunction with PCR to reliably detect Staphylococcus and Listeria spp..
DNA Sequencing
DNA sequencing technology has made it possible for scientists to sequence complete microbial genomes worldwide rapidly and effectively. New strategies offer the analysis and understanding of microbes at the molecular level in order to access the entire DNA sequences of microbial genomes. In response to the pathogenic invasion, researchers are attempting to identify pathogens in biological tissues and study variations in gene expression. These assist in the development of novel approaches for microbial pathogen detection and drug design and drug development. Each species of pathogen has a unique genomic signature that distinguishes itself from other organisms. The development of this nucleic acid signature for each microorganism of interest for quick and specific detection is one of the greatest challenges in the scientific field.
Plasmid Identification
For the typing of diverse species of Gram-negative and Gram-positive bacteria, plasmid DNA profiling has been utilized. It is adapted and used as an indicator in the field of health care to compare strains and evaluate the potential spread of a resistance gene. Interestingly, plasmids have unique properties, such as being transferable by conjugation under selective pressure to another strain, but can be acquired or lost spontaneously during the process. A misunderstanding of genetic relatedness to the isolate is generated by the gain or loss of plasmid and limits short-term epidemiological studies. For organisms like Enterobacteria and Staphylococcus spp., this is particularly useful.
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