Development of a novel dual priming oligonucleotide system‐based PCR assay for specific detection of Salmonella from food samples

Abstract

In this study, a novel dual priming oligonucleotide (DPO) system‐based polymerase chain reaction (PCR; DPO system‐based PCR) assay, which detected the fimY gene of Salmonella, was developed for the fast food testing. The DPO system‐based PCR assay allowed a wide range of annealing temperatures at 48–68°C to efficiently amplify fimY gene with an analytical detection limit of 1.2 × 102 CFU/ml for Salmonella in pure cultures and artificially contaminated food matrix. Significantly, the presence of a bubble‐like polydeoxyinosine (polyI) linker in the DPO system brought an unparalleled high specificity in the identification of target bacteria, and consequently, the false positives and mismatches of PCR process can be eliminated in priming. Applying the DPO system‐based PCR assay to 285 collected food samples revealed that 29 samples were positive in this assay, in accordance with the results of conventional culture‐based method, indicating a potential diagnostic capability. The high specificity of the DPO system‐based PCR indicates its great potential to be a quick, reliable and practical method for the detection of Salmonella in foods.

QCM Sensor Arrays, Electroanalytical Techniques and NIR Spectroscopy Coupled to Multivariate Analysis for Quality Assessment of Food Products, Raw Materials, Ingredients and Foodborne Pathogen Detection: Challenges and Breakthroughs

“…The practical utility of PCR for accurate detection of adulterated meat in mutton [27], detection of pork in meat and meat products [28], chicken adulteration [29], and the content of camel milk in adulterated milk samples was successfully demonstrated [30]. Importantly, the capability of PCR for the detection of a variety of foodborne pathogens, including Salmonella enterica, Escherichia Coli, and Campylobacter jejuni [31][32][33][34], Staphylococcus aureus [35], Toxoplasma gondii [36], and many foodborne pathogens in food samples have been recently reported. Moreover, the use of PCR for the GMO screening of food products with high specificity, good accuracy, and a low limit of detection has been demonstrated [37].…”

Visualization of copper nanoclusters for SARS-CoV-2 Delta variant detection based on rational primers design

“…A DPO primer is comprised of two separate priming regions joined by a polydeoxyinosine linker [ 22 ]. Since its discovery in 2007, it has been widely used in the detection of multiple viruses, including those with single nucleotide polymorphism (SNP), with high specificity and sensitivity [ 23 , 24 ].…”