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Simplified detection of the species of origin of antler velvets using single-stranded tag hybridization chromatographic printed-array strip

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Abstract

In this study, we developed a convenient and easy-to-use origin identification method for antler velvets based on a simple DNA extraction technique and single-stranded tag hybridization chromatographic printed-array strip (STH-PAS). The primer sets used to detect Cervus elaphus, Rangifer tarandus, and 12S rRNA did not engage in non-specific reactions such as primer dimer formation. In both the triplex and singleplex assays, the sensitivity was < 1 ng DNA. Moreover, Cervus elaphus DNA could be detected in OTC crude drug products. Although the detection sensitivity resulting from the simplified extraction was slightly lower than that obtained with extraction by conventional methods, the amount of DNA was sufficient even from a small sample. The choice of a triplex or singleplex assay will depend on the purpose of the test. For example, if it is important to determine whether the antler velvet is derived from Cervus elaphus or Rangifer tarandus, a triplex assay is appropriate. If it is necessary to explore whether antler velvet from Cervus elaphus is included in an OTC crude drug product, a singleplex assay using the Cervus elaphus primer set is informative. If it is necessary to explore whether powdered antler velvet includes counterfeit products (from Rangifer tarandus), a singleplex assay employing the Rangifer tarandus primer is appropriate. The singleplex assay detects minor components even at a 1,000:1 ratio. Our study thus demonstrated the utility of a method combining simple DNA extraction with STH-PAS for efficient identification of the origin of antler velvets.

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References

  1. Namba T (1980) Gensyoku-Wakanyaku-Zukan. Hoikusya, Osaka, Japan, pp279–283 (In Japanese)

  2. The Japanese standards for non-Pharmacopoeial crude drugs 2022 (2022) The Ministry of Health, Labor and Welfare

  3. Tokumoto H, Goto Y, Tsujimoto T, Maruyama T, Hakamatsuka T, Shiratori M, Arai R, Yamamoto Y, Yamada S, Okamoto T, Uchiyama N (2021) Discrimination method for crude drug antler velvet with microscopic examination and fluorescence analysis. Shoyakugaku zasshi 75:63–75. https://doi.org/10.24684/jspharm.75.2_63

    Article  Google Scholar 

  4. Yang F, Ding F, Chen H, He M, Zhu S, Ma X, Jiang L, Li H (2018) DNA Barcoding for the Identification and Authentication of Animal Species in Traditional Medicine. Evid Based Complement Alternat Med 1:5160254 Doi: https://doi.org/10.1155/2018/5160254

  5. Nakanishi H, Yoneyama K, Hayashizaki Y, Hara M, Takada A, Saito K (2019) Establishment of widely applicable DNA extraction methods to identify the origins of crude drugs derived from animals using molecular techniques. J Nat Med 73:173–178. https://doi.org/10.1007/s11418-018-1261-3

    Article  CAS  PubMed  Google Scholar 

  6. Hieno A, Li M, Afandi A, Otsubo K, Suga H, Kageyama K (2019) Rapid detection of Phytophthora nicotianae by simple DNA extraction and real-time loop-mediated isothermal amplification assay. J Phytopathol 167:174–184. https://doi.org/10.1111/jph.12785

    Article  CAS  Google Scholar 

  7. Yamamoto M, Alshahni MM, Tamura T, Satoh K, Iguchi S, Kikuchi K, Mimaki M, Makimura K (2018) Rapid detection of candida auris based on loop-mediated isothermal amplification (LAMP). J Clin Microbiol 56:e00591-e618. https://doi.org/10.1128/JCM.00591-18

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. https://www.t-bioarray.com/en_contents/technology.html

  9. Takarada Y, Kodera T, Kobayashi K, Nakajima C, Kawase M, Suzuki Y (2020) Rapid detection of rifampicin-resistant Mycobacterium tuberculosis, based on isothermal DNA amplification and DNA chromatography. J Microbiol Methods 177:106062. https://doi.org/10.1016/j.mimet.2020.106062

    Article  CAS  PubMed  Google Scholar 

  10. Koiwai K, Kodera T, Thawonsuwan J, Riani S, Kawase M, Kondo H, Hirono I (2018) Rapid diagnosis of three shrimp RNA viruses using RT-PCR-DNA chromatography. J Fish Dis 41:1309–1312. https://doi.org/10.1111/jfd.12821

    Article  Google Scholar 

  11. Monden Y, Takasaki K, Futo S, Niwa K, Kawase M, Akitake H, Tahara M (2014) A rapid and enhanced DNA detection method for crop cultivar discrimination. J Biotechnol 185:57–62. https://doi.org/10.1016/j.jbiotec.2014.06.013

    Article  CAS  PubMed  Google Scholar 

  12. Riztyan R, Takasaki K, Yamakoshi Y, Futo S (2018) Single-Laboratory validation of rapid and easy DNA strip for porcine DNA detection in beef meatballs. J AOAC Int 101:1653–1656. https://doi.org/10.5740/jaoacint.17-0394

    Article  CAS  Google Scholar 

  13. Tian L, Sato T, Niwa K, Kawase M, Tanner A, Takahashi N (2014) Rapid and sensitive PCR-dipstick DNA chromatography for multiplex analysis of the oral microbiota. Biomed Res Int. Doi: https://doi.org/10.1155/2014/180323.

  14. Kim M, Lee Y, Suh S, Kim H (2020) Species Identification of Red Deer (Cervus elaphus), Roe Deer (Capreolus capreolus), and Water Deer (Hydropotes inermis) Using Capillary Electrophoresis-Based Multiplex PCR. Foods 23:982. https://doi.org/10.3390/foods9080982

    Article  CAS  Google Scholar 

  15. Mitani T, Akane A (2008) Identification of animal species using the partial mitochondrial rRNA sequences. DNA Polymorphism 16:32–35 ((In Japanese))

    CAS  Google Scholar 

  16. Tian L, Sato T, Niwa K, Kawase M, Mayanagi G, Washio J, Takahashi N (2016) PCR-dipstick DNA chromatography for profiling of a subgroup of caries-associated bacterial species in plaque from healthy coronal surfaces and periodontal pockets. Biomed Res 37:29–36. https://doi.org/10.2220/biomedres.37.29

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We are grateful to Dr. Yutaka Yamamoto (Tochimoto Tenkaido) for providing some animal-derived crude drugs. We thank Dr. Mitsuo Kawase and Dr. Takuya Kodera (TBA Co., Ltd.) for technical assistance.

Funding

This research received no specific grants from any funding agency.

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Authors and Affiliations

  1. Department of Forensic Medicine, Juntendo University School of Medicine, 2-1-1, Hongo, Bunkyo-Ku, Tokyo, 113-8421, Japan

    Hiroaki Nakanishi, Kentaro Sakai & Kazuyuki Saito

  2. Department of Forensic Medicine, Saitama Medical University, 38 Morohongo, Moroyama, Saitama, 350-0495, Japan

    Hiroaki Nakanishi, Aya Takada, Katsumi Yoneyama & Kazuyuki Saito

  3. Tokyo Medical Examiner’s Office, Tokyo Metropolitan Government, 4-21-18, Otsuka, Bunkyo-Ku, Tokyo, 112-0012, Japan

    Aya Takada, Kentaro Sakai & Kazuyuki Saito

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  1. Hiroaki Nakanishi
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  2. Aya Takada
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  3. Katsumi Yoneyama
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  4. Kentaro Sakai
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  5. Kazuyuki Saito
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Contributions

Conceptualization, investigation, and writing, HN; review and editing, AT, KY, K. Sakai, and K. Saito. All authors have read and approved the published version of the manuscript.

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Correspondence to Hiroaki Nakanishi.

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Nakanishi, H., Takada, A., Yoneyama, K. et al. Simplified detection of the species of origin of antler velvets using single-stranded tag hybridization chromatographic printed-array strip. J Nat Med 77, 998–1004 (2023). https://doi.org/10.1007/s11418-023-01734-6

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  • DOI: https://doi.org/10.1007/s11418-023-01734-6

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