title: Advances and Challenges in Viability Detection of Foodborne Pathogens.
abstract: Foodborne outbreaks are a serious public health and food safety concern worldwide. There is a great demand for rapid, sensitive, specific, and accurate methods to detect microbial pathogens in foods. Conventional methods based on cultivation of pathogens have been the gold standard protocols; however, they take up to a week to complete. Molecular assays such as polymerase chain reaction (PCR), sequencing, microarray technologies have been widely used in detection of foodborne pathogens. Among molecular assays, PCR technology [conventional and real-time PCR (qPCR)] is most commonly used in the foodborne pathogen detection because of its high sensitivity and specificity. However, a major drawback of PCR is its inability to differentiate the DNA from dead and viable cells, and this is a critical factor for the food industry, regulatory agencies and the consumer. To remedy this shortcoming, researchers have used biological dyes such as ethidium monoazide and propidium monoazide (PMA) to pretreat samples before DNA extraction to intercalate the DNA of dead cells in food samples, and then proceed with regular DNA preparation and qPCR. By combining PMA treatment with qPCR (PMA-qPCR), scientists have applied this technology to detect viable cells of various bacterial pathogens in foods. The incorporation of PMA into PCR-based assays for viability detection of pathogens in foods has increased significantly in the last decade. On the other hand, some downsides with this approach have been noted, particularly to achieve complete suppression of signal of DNA from the dead cells present in some particular food matrix. Nowadays, there is a tendency of more and more researchers adapting this approach for viability detection; and a few commercial kits based on PMA are available in the market. As time goes on, more scientists apply this approach to a broader range of pathogen detections, this viability approach (PMA or other chemicals such as platinum compound) may eventually become a common methodology for the rapid, sensitive, and accurate detection of foodborne pathogens. In this review, we summarize the development in the field including progress and challenges and give our perspective in this area.
authors.affiliation: College of Veterinary Medicine, Nanjing Agricultural University Nanjing, China.
authors.forename: Dexin
authors.initials: D
authors.lastname: Zeng
authors.affiliation: College of Veterinary Medicine, Nanjing Agricultural UniversityNanjing, China; Animal Quarantine Laboratory, Jiangsu Entry-Exit Inspection and Quarantine BureauNanjing, China.
authors.forename: Zi
authors.initials: Z
authors.lastname: Chen
authors.affiliation: Animal Quarantine Laboratory, Jiangsu Entry-Exit Inspection and Quarantine BureauNanjing, China; Shanghai Entry-Exit Inspection and Quarantine BureauShanghai, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: College of Veterinary Medicine, Nanjing Agricultural University Nanjing, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Xue
authors.affiliation: Division of Molecular Biology, Center for Food Safety and Applied Nutrition, United States Food and Drug Administration, Laurel MD, USA.
authors.forename: Baoguang
authors.initials: B
authors.lastname: Li
keywords: PMA-qPCR; ethidium monoazide; false positive detection; foodborne pathogens; outbreaks; propidium monoazide; viability detection