Development of a PCR-based lateral flow strip assay for the simple, rapid, and accurate detection of pork in meat and meat products.

ABSTRACT

In recent years, adulteration of meat and meat products has become a major food safety issue. PCR and real-time PCR technologies are mainstream methods used to identify animal-derived components. However, these technologies rely highly on costly equipment and professional technicians; they are therefore difficult to use in resource-limited settings. In this study, a novel, highly sensitive molecular assay, Pig-PCR-Strip (Pig specific polymerase chain reaction-Lateral flow strip), was developed for rapid detection of pig and swine-derived components. The assay is based on PCR amplification, hybridization of the PCR product to the probe, followed by detection using a strip format. Using this format, the PCR product can be detected by the naked eye within 3 min, and provides a basis for the migration of species-specific detection to a point-of-care (POC) microfluidic format. The Pig-PCR-Strip can detect pork components at a concentration of 0.01% in adulterated meat, and the limit of detection is up to 10 fg of target DNA. The assay was specific to pork and did not cross-react with other non-target species. It also can be used for commercial samples and complex food samples. It is a promising new tool for detection of pig-derived meat and can be rapidly modified for identifying other species. It could be widely used for solving problems related to meat quality assurance, species authentication, and traceability.