Development of the Species-Specific Multiplex PCR and DNA Sequencing Methods for Rapid Authentication of Isatidis Folium and its Adulterants

Abstract

Isatis indigotica Fort. (family Cruciferae), is an herb widely used in traditional herbal medicine and its dried leave was named “ISATIDIS FOLIUM”. Baphicacanthus cusia (Ness) Bremek. and Polygonum tinctorium Ait. are commonly misused as ISATIDIS FOLIUM in Chinese Medicine pharmacy. For the purpose of being not misused, specific primers based on the sequence difference of chloroplast trnH-psbA intergenic spacer were designed and multiplex polymerase chain reaction method (multiplex PCR) was developed. In this study, 29 original herbal materials were analyzed and our results show that DNA size after multiplex PCR was able to distinguish variations between three herbs. DNA fragments of 464, 297, 170 base pairs (bps) were represented for I. indigotica and B. cusia and P. tinctorium , respectively. In conclusion, our investigations demonstrate that molecular identification method provides more accurate results for medicinal plants detection and good quality control of ISATIDIS FOLIUM.

Advancing Herbal Product Authentication: A Comprehensive Review Of DNA-Based Approach For Quality Control And Safety Assurance

“…The criteria considered, such as high copy number of chloroplast DNA, short amplicon length, specificity, and sensitivity, have significantly contributed to the effectiveness of these primers in accurately identifying and differentiating the target botanicals within herbal products. In the context of lower taxonomic level identification, the utilization of Species-specific Primers derived from the Internal Transcribed Spacer (ITS) region has proven to be a more effective alternative for detections (Hsieh et al, 2021;Sharma et al, 2017). British pharmacies have been pioneers in adopting trnH-psbA-derived species-specific primers for the authentication of Ocimum tenuiflorum (Cartwright, 2016) (Hsieh et al, 2021).…”

“…In the context of lower taxonomic level identification, the utilization of Species-specific Primers derived from the Internal Transcribed Spacer (ITS) region has proven to be a more effective alternative for detections (Hsieh et al, 2021;Sharma et al, 2017). British pharmacies have been pioneers in adopting trnH-psbA-derived species-specific primers for the authentication of Ocimum tenuiflorum (Cartwright, 2016) (Hsieh et al, 2021). These investigations contribute to the expanding scientific knowledge and emphasize the critical importance of rigorous authentication protocols in maintaining the quality control of herbal products.…”