期刊
MICROCHIMICA ACTA
卷 186, 期 9, 页码 -出版社
SPRINGER WIEN
DOI: 10.1007/s00604-019-3738-5
关键词
Species identification; ATPase subunit 8 gene; DNA extraction; Species-specific PCR; cytochrome b
资金
- CAMS Innovation Fund for Medical Sciences [2017-I2M-1-012]
- National Natural Science Foundation of China [31870059, 81760778]
- Shandong Natural Science Foundation [CR2014CL010]
- Drug Innovation Major Project [2018ZX09711001-007-003, 2017ZX09101002-003-003]
A PCR method is described to identify the species origin of various animal and human tissue-derived biochemical drugs. Four commercialized drugs, including spermary tablets, compound embryonic bovine liver extract tablets, spleen aminopeptide solution, and placenta polypeptide injection, were used as a proof-of-principle in this study. Primers were designed to amplify conservative regions of mitochondrial cytochrome b and ATPase 8 genes from beef, pork, lamb and human DNA, respectively. The specificity of primers for ATPase 8 gene is found to be higher than those for cytochrome b under the given experimental conditions. The amplicon sizes of ATPase 8 were 212, 271, 293 and 405 bp for pork, beef, lamb and human tissue, respectively. The minimum detectable concentration of DNA sample for species identification is 0.05-0.5 pg center dot mu L-1. The species origin can be distinguished by this method in extremely low concentrations of template DNAs extracted. Conceivably, this PCR method for meat authentication may be extended to quality control of other biochemical drugs and raw materials.
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