The Design of Specific Primers of Pheretima aspergillum<\i> and the Identification of Adulterants

OBJECTIVE The specific primers were designed to identify Pheretima aspergillum and its adulterants. METHODS The DNA of Pheretima aspergillum and its adulterants were extracted and amplified using the COⅠsequences as primers. The amplification products were sequenced afterwards. By comparing the COⅠsequences of Pheretima aspergillum with its adulterants， the variation sites were found in Pheretima aspergillum and the specific primers were designed to identify Pheretima aspergillum and its adulterants. PCR reaction system was optimized. RESULTS 750 bp fragments could be detected in the products of other animal medicinal materials using the COⅠsequences as primers， which meant the primers were not specific enough to distinguish the Pheretima aspergillum from its adulterants. Based on the COⅠsequences of amplification products， the specific primers PA-f/PA-r were designed. Through the established PCR reaction system， 574 bp fragments were amplified from DNA templates of Pheretima aspergillum. All the adulterants had no bands. CONCLUSION The designed primers sequences could accurately and successfully distinguish the Pheretima aspergillum from its adulterants， providing an effective method for the identification of Pheretima aspergillum.