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Development of species identification tests targeting the 16S ribosomal RNA coding region in mitochondrial DNA

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Abstract

This report describes the development of a species testing system based on the diversity of nucleotide sequences in mitochondrial DNA (mtDNA) among species. Five species, human, cow, pig, dog, and cat, were considered. The partial nucleotide sequences in 16S ribosomal RNA coding region were chosen as the target for discriminating the species. The sequence diversities of this approximately 400 bp long region ranged from 15.7 to 24.1% among the five species. Sequencing analysis of this target on 50 individuals of each species (53 for dogs) revealed that the nucleotide sequences were well preserved within species. Species-specific PCR for each species was also designed, and satisfactory results with regard to both sensitivity and specificity were obtained. A validation study with DNA extracted from bovine bone exposed to the environment revealed that the PCRs designed in this study worked correctly. From the results obtained, it was shown that this testing system could be a good tool for species identification. One successful case report is also demonstrated.

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Acknowledgments

The authors wish to thank Dr. Shinji Kaneko and Dr. Hiroe Kaneko, veterinary surgeons of Kaneko animal hospital, Dr. Jyn Ayai and Dr. Chiharu Taniguchi, veterinary surgeons of Ayai animal hospital, Dr. Tomoko Inoue, veterinary surgeon of Heiwa animal hospital, and Dr. Kazuhide Morimoto, veterinary surgeon of Hiroshima-higashi livestock hygene service center, for their kind donation of samples. The authors also thank two anonymous reviewers for their helpful comments. This study was supported by the research fund of the National Research Institute of Police Science.

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Authors and Affiliations

  1. National Research Institute of Police Science, 6-3-1, Kashiwanoha, Kashiwa, Chiba, 277-0882, Japan

    Kazuhiko Imaizumi, Tomoko Akutsu, Sachio Miyasaka & Mineo Yoshino

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  1. Kazuhiko Imaizumi
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  2. Tomoko Akutsu
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  3. Sachio Miyasaka
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  4. Mineo Yoshino
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Correspondence to Kazuhiko Imaizumi.

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Imaizumi, K., Akutsu, T., Miyasaka, S. et al. Development of species identification tests targeting the 16S ribosomal RNA coding region in mitochondrial DNA. Int J Legal Med 121, 184–191 (2007). https://doi.org/10.1007/s00414-006-0127-5

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  • DOI: https://doi.org/10.1007/s00414-006-0127-5

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