OBJECTIVE To establish a rapid method for distinguishing Aucklandia costus, Dolomiaea Souliei and Inula racemosa by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).
METHODS The ITS gene sequences of Aucklandia costus, Dolomiaea souliei and Inula racemosa were compared, restriction enzymes were screened and identification primers were designed. The PCR amplification and digestion reaction conditions were optimized, and the methodological examinations and statistical analysis were carried out.
RESULTS When the annealing temperature was 60 ℃ and the cycle number was 30, a single DNA band of 500~750 bp could be detected. When the substrate was 10 μL and the digestion time was 15 min, BsaHI enzyme could cut Aucklandia costus, Dolomiaea souliei and Inula racemosa into DNA bands of different sizes in a certain range. The results of methodology and statistics were good.
CONCLUSION The PCR-RFLP method established in this study can accurately identify the three medicinal materials of Aucklandia costus, Dolomiaea souliei and Inula racemosa, avoid the mixed use of these three medicinal materials, and ensure the safety of the clinical medication.
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