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Microchimica Acta

, 186:634 | Cite as

Specific PCR method for detection of species origin in biochemical drugs via primers for the ATPase 8 gene by electrophoresis

  • Limei Ai
  • Juanjuan Liu
  • Yu Jiang
  • Weiwei Guo
  • Ping WeiEmail author
  • Liping BaiEmail author
Original Paper

Abstract

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·μL−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.

Graphical abstract

A specific PCR method was developed for the detection of species origin in biochemical drugs via species-specific primers targeting mitochondrial ATPase 8 genes. The PCR products were separated by gel electrophoresis and species origins were indicated by comparison to references.

Keywords

Species identification ATPase subunit 8 gene DNA extraction Species-specific PCR cytochrome b 

Notes

Acknowledgements

This research was supported by grants from CAMS Innovation Fund for Medical Sciences (2017-I2M-1-012), National Natural Science Foundation of China (31870059 & 81760778), Shandong Natural Science Foundation (CR2014CL010) and The Drug Innovation Major Project (2018ZX09711001-007-003 & 2017ZX09101002-003-003). We specially thank Dr. Mengxin Geng in Texas A&M University for critical reading of the manuscript.

Compliance with ethical standards

The author(s) declare that they have no competing interests.

Supplementary material

604_2019_3738_MOESM1_ESM.docx (595 kb)
ESM 1 (DOCX 595 kb)

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Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2019

Authors and Affiliations

  1. 1.NHC Key Laboratory of Biotechnology of Antibiotics, Institute of Medicinal BiotechnologyChinese Academy of Medical Sciences & Peking Union Medical CollegeBeijingChina
  2. 2.Department of medical immunology, School of Basic Medical SciencesTaishan Medical UniversityTaianChina
  3. 3.CAMS Key Laboratory of Synthetic Biology for Drug Innovation, Institute of Medicinal BiotechnologyChinese Academy of Medical Sciences & Peking Union Medical CollegeBeijingChina

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