Authentication of medicinal herbs using PCR-amplified ITS2 with specific primers.
Different parts of medicinal herbs have long been used as traditional Chinese drugs for treating many diseases, whereas materials of similar morphology and chemical fingerprints are often misidentified. Analyses of sequence variations in the nuclear ribosomal DNA (rDNA) internal transcribed spacer (ITS) have become a valid method for authentication of medicinal herbs at the intergenic and interspecific levels. DNA extracted from processed materials is usually severely degraded or contaminated by microorganisms, thus generates no or unexpected PCR products. The goal of this study is to apply the ITS fragments selectively amplified with two designed primer sets for efficient and precise authentication of medicinal herbs. The designed primers led to an accurate PCR product of the specific region in ITS2, which was confirmed with DNA extracted from 55 processed medicinal herbs belonging to 48 families. Moreover, the selectively amplified ITS2 authenticated five sets of easily confusable Chinese herbal materials. The designed primers were proven to be suitable for a broad application in the authentication of herbal materials.
Chiou
Shu-Jiau
SJ
Department of Life Science and Institute of Bioinformatics and Structural Biology, National Tsing Hua University, Hsinchu, Taiwan, ROC.
Yen
Jui-Hung
JH
Fang
Cheng-Li
CL
Chen
Hui-Ling
HL
Lin
Tsai-Yun
TY
D017931:DNA Primers; D018744:DNA, Plant / Q000032:analysis*; D021903:DNA, Ribosomal Spacer / Q000032:analysis*; D004365:Drugs, Chinese Herbal / Q000032:analysis*; D006801:Humans; D008517:Phytotherapy / Q000592:standards*; D010946:Plants, Medicinal / Q000235:genetics*; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D018744:DNA, Plant; D021903:DNA, Ribosomal Spacer; D004365:Drugs, Chinese Herbal