Development of deft amplification refractory mutation sequencing system (ARMSS) for discriminating Pilos antler based on a short cytochrome b (Cytb) gene

Abstract Pilos antler (Lu-Rong in Chinese) is a famous traditional medicine in China. Many adulterants have been discovered in Chinese markets in recent years. However, few DNA-based methods are effective for discrimination of this DNA-degraded animal medicine. Here, novel and deft amplification refractory mutation sequencing system (ARMSS), integrating the advantages of the amplification refractory mutation system (ARMS) and the short DNA barcode, was first developed to discriminate Pilos antler from its adulterants. We aimed to provide a new sight and inspiration for deft detection. The results showed that developed ARMS achieved strong specificity and high sensitivity in rapid identification, while the short Cytb gene was of excellent identification power in terms of accurate identification, which suggested that ARMSS successfully integrated the advantages of the ARMS and short DNA barcode, and that it was useful for deft detection. Our study determined that the deft ARMSS could be the well candidate for discrimination of Pilos antler, as well as be a valuable tool for deft identification of Chinese medicine.

[1]  Damon P. Little,et al.  Choosing and Using a Plant DNA Barcode , 2011, PloS one.

[2]  Li Zhang,et al.  Pilose antler polypeptides promote chondrocyte proliferation via the tyrosine kinase signaling pathway , 2011, Journal of occupational medicine and toxicology.

[3]  D Chen,et al.  [Character identification of 12 kinds of pilose antler medicinal materials]. , 1999, Zhong yao cai = Zhongyaocai = Journal of Chinese medicinal materials.

[4]  M. Nei,et al.  MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. , 2011, Molecular biology and evolution.

[5]  Ting-Guo Kang,et al.  [Microscopical identification and hierarchical cluster analysis of seven kinds of pilose antler velvet]. , 2009, Zhong yao cai = Zhongyaocai = Journal of Chinese medicinal materials.

[6]  Xiao‐he Xiao,et al.  Forensic DNA Barcoding and Bio-Response Studies of Animal Horn Products Used in Traditional Medicine , 2013, PloS one.

[7]  Ye Dj Microscopic identification of pilose antler and deerhorn , 1986 .

[8]  I. Day,et al.  An efficient procedure for genotyping single nucleotide polymorphisms. , 2001, Nucleic acids research.

[9]  Yong-zhong Wang,et al.  Detection of hepatitis B virus A1762T/G1764A mutant by amplification refractory mutation system , 2014, The Brazilian journal of infectious diseases : an official publication of the Brazilian Society of Infectious Diseases.

[10]  P. Ong,et al.  DNA barcoding of fishes of Laguna de Bay, Philippines , 2011, Mitochondrial DNA.

[11]  P. Gupta,et al.  Sequence characterization and polymerase chain reaction–restriction fragment length polymorphism of the mitochondrial DNA 12S rRNA gene provides a method for species identification of Indian deer , 2008, Mitochondrial DNA.

[12]  Ying Liu,et al.  Authentication of official Da-huang by sequencing and multiplex allele-specific PCR of a short maturase K gene. , 2013, Genome.

[13]  Shilin Chen,et al.  The Short ITS2 Sequence Serves as an Efficient Taxonomic Sequence Tag in Comparison with the Full-Length ITS , 2013, BioMed research international.