Developing a Multiplex PCR Technique for the Identification of Halal and Non-Halal Meats

Food safety is a major concern in most Muslim countries (e.g., Iran), where halal and non-halal meat adulteration is believed to cause significant social and health risks. Thus, developing a valid and reliable detection method to distinguish between halal and non-halal meats in processed foods is crucial, particularly for small-scale laboratories. In this regard, this current work aimed at designing and optimizing species-specific primers to identify halal and non-halal meats in processed foods with the use of the NADH Dehydrogenase and ATP Synthase genes via simplex and multiplex PCR assays. The results depicted that the DNA extracted from processed beef, poultry, and pork could be effectively amplified using the NADH Dehydrogenase and ATP Synthase primers, producing amplicon bands that were visible and consistent with the expected size in both simplex and multiplex PCR experiments. Despite the fact that non-specific bands were observed in several primer sets, the primary target bands remained distinct and easily identifiable. The existence of such primers is expected to promote the efficiency of halal food authentication, particularly in smallscale laboratories in rural regions of Iran.