Continuing the discussion, we often get questions on investigating the quality of a method and the possibility of combining it with a company’s own internal routine testing, both for simplicity and defensible results. I often get questions like “is the assay ready to use?”, “is the method repeatable and sensitive enough for religious purposes?”, or “are there multiplex assays for multiple targets?”
This last question in particular requires a complex answer. Multiplex assays like this are quite risky. For example, there must be no sequence homology between the various primers. The length of the various amplicons must be different to discriminate between them, but also fall within a certain range. And finally, the pairs of primers used must be sufficient to amplify all the loci of a gene or a region of interest, otherwise one primer can cannibalise the other and produce a false negative result.
MC: Could you describe some real-world scenarios where meat speciation testing played a crucial role in addressing customer needs or challenges? What were the outcomes or benefits observed?
GT: Some time ago in Italy there was a contamination of horse meat in stuffed pasta not declared on the label, and this led to quite the scandal across Europe. These could have been avoided with stringent analytical protocol and molecular tests.
MC: Bio-Rad recently launched a new line of PCR-based speciation kits called ID-Check. Can you provide more details about these kits?
GT: Bio-Rad, already an industry leader for PCR solutions for food pathogen testing, recently launched the ID-Check Solution for eight different meat species (bovine, turkey, duck, goat, chicken, horse, swine and sheep), plus an extraction kit in a ready‑to‑use format that is easy to use, with results in less than two hours that deliver reliable and reproducible results with high sensitivity (limit of detection (LOD) at least 0.01%) and specificity (100%). After sample preparation and optimised DNA extraction, results are ready after a 75-minute PCR run detecting species‑specific mitochondrial DNA with proprietary primers and probes.
MC: Can you explain how DNA‑based methods like ID‑Check are used for meat speciation, and what advantages they offer over traditional techniques?
GT: The methodologies used today in species determination are mainly based on DNA and proteins/peptides and other less used techniques such as triacylglycerol. The PCR technique certainly has many advantages over an immunological ELISA test, given that the latter can suffer from issues with cross-reactivity, detection limit and given that antibody stability can be sensitive to environmental conditions such as temperature and pH.
MC: What challenges might arise when using a DNA-based method such as ID-Check in processed or highly cooked foods? How does the ID-Check solution help overcome these challenges?
GT: The protein subjected to cooked temperatures can degrade or lose activity over time, compromising the reliability and reproducibility of the test. Without an internal reaction control you are blind to possible false negative results, while the Bio-Rad ID‑Check Speciation Kit detects mitochondrial DNA ensuring a low LOD compared even to single copy genomic DNA targets and the possibility to determinate a high mutation rate. In addition, each reaction contains an internal amplification control.
MC: How can the ID-Check Solution be used to help companies ensure they are getting accurate results they can trust?
GT: ID-Check Speciation kits are a premium quality PCR solution that provide general laboratory and research personnel with an easy and ready-to-use kit for the detection of DNA from select meat species. The kits target mitochondrial DNA in food, feed and environmental samples.
Our solution offers quality, linked to the brand, high performance, data security for all types of laboratories, including service labs, in-house testing labs, R&D labs and regulatory authorities to help trade and enable consumers to make informed choices.
Conclusion
By embracing transparent practices, we can hope to establish a more resilient, ethical and safe food system for future generations. We can put an end to meat fraud and foster a culture of accountability at all levels of the food supply chain.
As technology continues to advance and the use of molecular methods becomes the standard for accuracy and sensitivity, Bio‑Rad’s ID-Check Speciation Solution gives users the ability to optimise their vulnerability assessment and critical control point plans and work towards vigilance and continuous improvement.
For more information on Bio-Rad’s ID-Check Speciation Solution, please visit:
www.bio-rad.com/speciation