Objective: To establish a method for rapid identification of Liriopes Radix adulterated in Ophiopogon Radix medicinal materials and decoction pieces by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Methods: By comparison ITS sequences restriction site of Ophiopogonis Radix and Liriopes Radix, the specific restriction site of Liriopes Radix were selected, and the primers for PCR-RFLP reaction were designed. The different Ophiopogonis Radix and Liriopes Radix were amplified by PCR, PCR-RFLP reaction system annealing temperature, primer concentration, cycle number, enzyme digestion reaction time were and restriction endonuclease type optimized, and accuracy of the method were investigated. In addition, the PCR-RFLP method which had established was used to investigate the applicability of Ophiopogonis Radix, Liriopes Radix and adulterated sample from different sources. Results: A PCR-RFLP method for identifying Ophiopogonis Radix mixed with Liriopes Radix was established with the annealing temperature at 58 ℃ and the number of cycles of 30. Liriopes Radix and Liriopes Radix adulterated in Ophiopogonis Radix could be digested by PmlI restriction endonuclease after amplification with specific primers, the two single DNA bands were detected between 100-250 bp, which were not found in Ophiopogonis Radix. This method could be used to identify Ophiopogonis Radix, Liriopes Radix and the adulterated sample, and the minimum detection limit of this method for Liriopes Radix adulterated in Ophiopogonis Radix was 5%. Conclusion: The established PCR-RFLP method is specific and good stable, can be applied to the qualitatively identify Ophiopogonis Radix, Liriopes Radix and its adulterated sample, which can be used to establish supplem entary method.
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