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

Plants belonging to the Hoodia 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 Hoodia in commercial products. These factors highlight the need for a reliable genus-level detection method for Hoodia to support the control of illegal trade and the verification of marketed Hoodia -based products. To address this need, we developed a real-time PCR method specific to Hoodia 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 Hoodia 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 Hoodia monitoring and authentication in commercial products. • A real-time PCR method was developed and validated for Hoodia genus detection. • 13 new Hoodia ITS2 sequences were generated in-house to optimize assay design. • The assay specifically detected Hoodia with no false positives or negatives. • The highly sensitive assay detected fewer than 25 DNA copies. • The assay enabled Hoodia monitoring in commercial herbal supplements.