Establishment of a Rapid PCR Detection Method for Antrodia salmonea and A. cinnamomea

摘要

Both Antrodia cinnamomea and A. salmonea are fungi of the Polyporaceae. Antrodia cinnamomea, which only grows on the inner side of the trunk of Cinnamomum kanehirai Hayata (Lau-raceae), is a precious medicinal fungus; A. salmonea causes brown heart rot disease of Cunningha-mia konishii Hayata (Cunninghamieae) in Taiwan. It is difficult to distinguish A. cinnamomea from A. salmonea because of their morphological similarities. Antrodia salmonea is commonly used as a counterfeit substitute for A cinnamomea and sold by dishonest merchants. In 2004, these 2 fiingi were first distinguished by the pore surface color of the basidiomata, host preferences, and mating types. However, accurate identification relies on professional training and experience. More-convenient and-persuasive methods are necessary for precisely identifying A. cinnamomea. In this study, we applied 6 fungal primer pairs in polymerase chain reaction (PCR) assays, and analyzed sequences of amplified DNA fragments between A. cinnamomea and A. salmonea. Results showed that one of these primer pairs could amplify a particular DNA fragment from A. salmonea, which was approximately 1.5 kb longer than that from A. cinnamomea. Based on this 1.5-kb difference in sequences, another primer pair named Acl-F/Acl-R was designed for the specific detection of A. salmonea. A specific fragment, of 219 bp, was yielded only from A. salmonea, whereas no fragment was yielded from A. cinnamomea by the PCR assay. In this study, we established a rapid and accurate identification technique for ^4. salmonea^ which will be helpful in rapidly differentiating A. salmonea from A. cinnamomea. Additionally it also saves significant time as the entire procedure only takes about 1 h and has high sensitivity. This identification method can provide objective evidence for industry and public institutions.