Molecular authentication of the animal crude drug Sailonggu (bone of Myospalax baileyi).
Two pairs of allele-specific diagnostic primers (SL1L/SL1H and SL2L/SL2H) for distinguishing the Chinese crude drug Sailonggu (bone of plateau zokor, Myospalax baileyi) from its substitutes were designed based on complete sequences of mitochondrial 12S rRNA and cytochrome b genes of the original animals of Myospalacinae, bamboo rat Rhizomys sinensis and black lipped pika Ochotona curzoniae. Total DNA was extracted from crude drug samples and original animals. Allele-specific diagnostic PCRs were performed using these primers with the total DNA as a template annealing at 65 degrees C. Positive amplifications were obtained from all DNA templates of Sailonggu and M. baileyi, whereas negative amplifications resulted from those of other zokors, the bamboo rat and black lipped pika. These results indicate that Sailonggu samples can be definitely distinguished from their substitutes by diagnostic PCR, and no incorrect discrimination was found under the same reaction conditions. Each of the two diagnostic primer pairs can be used to distinguish crude drug Sailonggu from its substitutes or adulterants. The three Sailonggu samples studied were diagnosed as genuine Sailonggu. In addition, the results of sequence alignment and phylogenetic analysis are congruent with that of the allele-specific diagnostic PCR.
Zhou
Caiquan
C
Jiangsu Key Lab for Bioresource Technology, College of Life Sciences, Nanjing Normal University, China.
Zhou
Kaiya
K
Zhang
Shaolai
S
D000483:Alleles; D000818:Animals; D001842:Bone and Bones / Q000737:chemistry*; D045304:Cytochromes c / Q000235:genetics; D017931:DNA Primers; D008516:Medicine, Chinese Traditional; D008928:Mitochondria / Q000737:chemistry; D004364:Pharmaceutical Preparations / Q000032:analysis*; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012377:Rodentia / Q000235:genetics*; D016415:Sequence Alignment
D017931:DNA Primers; D004364:Pharmaceutical Preparations; D045304:Cytochromes c