Real-time PCR to target Hoodia in herbal supplements: a tool for conservation and trade regulation

Plants belonging to the <i>Hoodia</i> genus are widely used as ingredients in herbal dietary supplements. Rising demand and overharvesting have led to supply shortages, adulteration, and CITES-regulated trade, threatening the authenticity and safety of <i>Hoodia</i> in commercial products. These factors highlight the need for a reliable genus-level detection method for <i>Hoodia</i> to support the control of illegal trade and the verification of marketed <i>Hoodia</i>-based products. To address this need, we developed a real-time PCR method specific to <i>Hoodia</i> genus targeting the ITS2 region, a molecular marker with high variability enabling detection and differentiation of plant species or genera. Due to the limited number of publicly available <i>Hoodia</i> ITS2 sequences, 13 additional sequences were generated in-house to support assay design. This real-time PCR method was validated in compliance with international standards, demonstrating high specificity, high sensitivity, and inter-laboratory transferability. Its applicability was verified by testing commercial herbal dietary supplements. This study provides enforcement laboratories with a validated molecular tool to support <i>Hoodia</i> monitoring and authentication in commercial products.