Polymerase chain reaction-restriction fragment length polymorphism test for the authentication of herbal Chinese medicines

A method for determining whether a given herbal or animal material is that of Codonopsis modesta, Codonopsis nervosa, Codonopsis pilosula, Codonopsis tangshen, Platycodon grandiflorus, Campanumoea javanica, Epimedium brevicornum, Epimedium koreanum, Epimedium pubescens, Epimedium wushanense, Tulipa edulis, or Pheretima aspergillus, which comprises:

• a) extracting rDNA from the herbal or animal material;
• b) amplifying an ITS1-5.8S-ITS2 region of the extracted rDNA using oligonucleotide primers whose nucleotide residue is conserved across the plant kingdom and which flank the ITS1-5.8S-ITS2 region of Codonopsis modesta, Codonopsis nervosa, Codonopsis pilosula, Codonopsis tangshen, Platycodon grandiflorus, Campanumoea javanica, Epimedium brevicornum, Epimedium koreanum, Epimedium pubescens, Epimedium wushanense, Tulipa edulis, and Pheretima aspergillus;
• c) digesting amplified nucleic acid with a restriction endonuclease so as to generate restriction fragments;
• d) separating the restriction fragments obtained in step c) to generate a restriction fragment length profile;
• e) comparing this restriction fragment length profile with known restriction fragment length profiles of herbs and animals, thereby determining whether the material is that of either Codonopsis modesta, Codonopsis nervosa, Codonopsis pilosula, Codonopsis tangshen, Platycodon grandiflorus, Campanumoea javanica, Epimedium brevicornum, Epimedium koreanum, Epimedium pubescens, Epimedium wushanense, Tulipa edulis, or Pheretima aspergillus or whether the herbal or animal material is from an entirely different source.