Work

Development of a MIRA-CRISPR/Cas12a-based nucleic acid detection system for the discrimination of Panax ginseng and Panax quinquefolium

Year: 2025

Type: article

Abstract: Accurate identification of closely related herbal species is essential for ensuring the safety, efficacy, and authenticity of traditional Chinese medicine (TCM) products. Panax ginseng (PG) and Panax quinquefolium (PQ) are two widely used ginseng species with similar morphological characteristics but distinct pharmacological profiles and market values. We developed a nucleic acid detection platform that integrates multienzyme isothermal rapid amplification (MIRA) with CRISPR/Cas12a-based fluorescence or lateral flow assay (LFA) for the rapid discrimination of PG and PQ. By targeting highly divergent mitochondrial gene regions, we designed species-specific crRNAs that enabled precise identification of PG and PQ with high sensitivity and specificity. By optimizing the conditions, the system can effectively distinguish PG and PQ, with a fluorescence detection limit of 10 -4 ng/μL and a LFA detection limit of 10 -3 ng/μL. It was also validated using 29 commercial samples including complex Chinese medicines. Compared with traditional DNA barcoding, the MIRA-CRISPR/Cas12a system demonstrated superior performance in detecting mixed or processed herbal products. This method offers a promising solution for point-of-care testing (POCT) in TCM quality control and provides a foundation for broader applications of CRISPR-based molecular authentication in herbal medicine.

Source: Journal of Ginseng Research

DOI: 10.1016/j.jgr.2025.11.011