Polymerase chain reaction with lateral flow sensor assay for the identification of horse meat in raw and processed meat products.

A simple, rapid and accurate detection method for the authentication of animal species is urgently required in the food detection field. The present study established a horse-specific polymerase chain reaction integrated with a lateral flow sensor assay (Horse-PCR-LFS) for the rapid detection of horse meat. In this test, a cytb gene sequence of horse was amplified using PCR, the PCR amplicon was checked with the lateral flow sensor assay, and the result of the sensor can be read within 2-3 min by the naked eye. The detection limit of the test was up to 0.01% horse meat in artificially adulterated meat mixtures, the assay also successfully detected horse DNA in various commercial food samples. As a rapid and user-friendly molecular detection tool, this test provides an accurate detection format for the identification of horse and offers solutions to problems related to animal meat adulteration and animal-origin food safety and traceability.

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

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.

Development of a single-tube nested PCR-lateral flow biosensor assay for rapid and accurate detection of Alternaria panax Whetz.

Alternaria panax Whetz causes one of the most commonly occurring and serious diseases in ginseng cultivation, and may cause significant production and economic losses in the ginseng industry. Rapid, early, and accurate identification of Alternaria panax Whetz is an essential prerequisite for the effective prevention and control of further infection spread. In this work, a rapid and accurate molecular diagnostic method, a single-tube nested PCR-lateral flow biosensor assay (STNPCR-LFBA), was developed for rapid identification of Alternaria panax Whetz. The STNPCR-LFBA was 100 times more sensitive than the traditional PCR-LFBA. Besides that, the PCR product was checked by a lateral flow biosensor assay, which provided a basis for the migration of the detection technology to a point-of-care test (POCT) format. STNPCR-LFBA was specific to Alternaria panax Whetz, and no cross-reactions were observed in other non-target samples; the limit of detection was up to 0.01 pg of Alternaria panax Whetz genomic DNA. STNPCR-LFBA could also be used for specific identification of Alternaria panax Whetz in real samples. STNPCR-LFBA is useful for identifying Alternaria panax Whetz due to its rapidity, accuracy, and simple manipulation.