Polymerase chain reaction - restriction fragment length polymorphism test for the authentication of traditional chinese medicines

Abstract

This invention provides a procedure for authentication of plant and animal materials used as traditional Chinese medicine is described. This method amplifies and detects the discrete and species-specific RFLP patterns in the region of rDNA. The present invention offers a reliable and definite way to identify morphologically similar Chinese medicine, using a minute amount of biological samples. Its application in the authentication of American and Oriental ginsengs is illustrated in detail.

What is claimed is:  
     
         1 . An isolated nucleic acid having the ITS1-5.8S-ITS2 sequence of  Codonopsis modesta.    
     
     
         2 . An isolated nucleic acid having the ITS1-5.8S-ITS2 sequence of  Codonopsis nervosa.    
     
     
         3 . An isolated nucleic acid having the ITS1-5.8S-ITS2 sequence of  Codonopsis pilosula.    
     
     
         4 . An isolated nucleic acid having the ITS1-5.8S-ITS2 sequence of  Codonopsis tangshen.    
     
     
         5 . An isolated nucleic acid having the ITS1-5.8S-ITS2 sequence of  Platycodon grandiflorus.    
     
     
         6 . An isolated nucleic acid having the ITS1-5.8S-ITS2 sequence of  Campanumoea javanica.    
     
     
         7 . An isolated nucleic acid having the ITS1-5.8S-ITS2 sequence of  Epimedium brevicornum.    
     
     
         8 . An isolated nucleic acid having the ITS1-5.8S-ITS2 sequence of  Epimedium koreanum.    
     
     
         9 . An isolated nucleic acid having the ITS1-5.8S-ITS2 sequence of  Epimedium pubescens.    
     
     
         10 . An isolated nucleic acid having the ITS1-5.8S-ITS2 sequence of  Epimedium wushanense.    
     
     
         11 . An isolated nucleic acid having the ITS1-5.8S-ITS2 sequence of  Tulipa edulis.    
     
     
         12 . An isolated nucleic acid having the ITS1-5.8S-ITS2 sequence of  Pheretima aspergillus.    
     
     
         13 . A method for authentication of a given herbal or animal material which comprises: 
 a) extracting DNA from the herbal or animal material;    b) amplifying an ITS1-5.8S-ITS2 region of the extracted DNA using oligonucleotide primers whose nucleotide sequences are conserved across the plant or animal kingdoms and flank the ITS1-5.8S-ITS2 region;    c) digesting the amplified nucleic acid with one or more restriction endonucleases so as to generate restriction fragments;    d) separating the restriction fragments obtained in step c) to generate a restriction fragment length profile;    e) comparing the restriction fragment length profile obtained in step d) with a database of known restriction fragment length profiles of herbs and animals so as to thereby determine whether the material is one of the herbs or animals in the database or is from an entirely different source, thereby authenticating a given herbal or animal material.