Development of a species-specific PCR-lateral flow assay integrated with SYBR Safe for rapid authentication of commercial Polygoni Multiflori Caulis (Ye-Jiao-Teng).

Pleuropterus multiflorus (PM) is a widely used medicinal plant in traditional Chinese medicine (TCM). However, the closely related species Pleuropterus angulatus (PA) is commonly misidentified as PM due to morphological similarities, leading to frequent adulteration in commercial Polygoni Multiflori Caulis (PMC) products. To address this issue, we developed a species-specific PCR-lateral flow assay integrated with SYBR Safe for rapid and species-specific authentication of PM and PA. Species-specific primers were designed based on a diagnostic single nucleotide polymorphism (SNP) within the chloroplast trnL-trnF region. After validation of specificity, visual detection of PM and PA was achieved using lateral flow strips. The assay demonstrated detection limits of 0.5 ng/μL for PM and 0.05 ng/μL for PA genomic DNA. The application of this assay to eighteen commercial PMC products revealed the presence of PA in all samples, with only one product containing both PM and PA, indicating widespread adulteration in the herbal marketplace. In conclusion, the developed PCR-lateral flow assay provides a rapid, sensitive, and user-friendly molecular diagnostic tool suitable for authenticity testing of herbal products and quality control during pharmaceutical manufacturing.